Diagram of a tank screening for dysentery. Laboratory diagnosis of dysentery (literature review)

Dysentery is a severe intestinal infection characterized by an acute onset. Microbiological diagnostics dysentery consists in isolating the pathogen from the fecal masses of the patient by sowing in a special nutrient medium. The disease must be differentiated from other intestinal diseases and poisonings. Early diagnosis and timely treatment will help to avoid complications.

The importance of timely diagnosis

Recognizing dysentery in practice is not so easy because there are infectious and non-infectious diseases with similar clinical manifestations. A characteristic feature of causative agents of dysentery (shigella) is the ability to change resistance to antibacterial drugs. An untimely diagnosed disease will lead to infection a large number of people. Misuse of antibiotics is the reason for the emergence of resistance in bacteria, leading to mass infections and epidemics with deaths. The source of infection is patients and carriers of bacteria that excrete pathogenic microorganisms with fecal matter. Incubation period dysentery - 2-3 days.

Clinical symptoms of the disease

• Sudden fever with a body temperature of 40 degrees or more.
• Diarrhea more than 10 times a day.
• The appearance in the stool of blood, mucus, in rare cases pus.
• Loss of appetite up to complete absence.
• Nausea and vomiting.
• Cutting in abdomen and right hypochondrium.
• Pain in the rectum.
• Dehydration.
• Dry tongue with white coating.
• Arrhythmia.
• Decreased blood pressure.
• Disorders of consciousness.

Diagnostic procedures

The doctor puts the diagnosis of dysentery only after the conducted researches.

Diagnosis of the disease includes generally accepted and special methods that establish not only final diagnosis, but also assessing the level of disorders of the digestive organs. With dysentery, the diagnosis is made on the basis of the epidemiological picture of the disease, clinical symptoms and conducted research. The main laboratory diagnostics is the analysis of feces for microbiology, sowing up to 80% of pathogens. The serological method is carried out no earlier than the 5th day of the disease, this type of study complements, but does not replace the microbiological analysis. Other methods:

• Coprological examination is a simple and affordable clinical method that detects mucus, blood streaks, erythrocytes, neutrophils (up to 50 per field of view) and altered epithelial cells.
• Sigmoidoscopy - allows you to monitor the healing process. Not applicable to children.
• Allergy test method - helper method, based on the taking of an allergic skin test with dysentery (Tsuverkalov's method).

General blood analysis

Immunity cells destroy dysentery pathogens even in the intestines, and severe cases of the disease occur when bacteria enter the lymph nodes, followed by entry into the bloodstream. A blood test for dysentery assesses the patient's condition and allows you to respond in time to possible complications. An increase in the erythrocyte sedimentation rate is a laboratory indicator that characterizes the degree of inflammation. Also, dysentery causes an increase in the concentration of stab neutrophils and monocytes.

How to donate feces for a coprogram?

To confirm the disease, a stool test is performed. Coprogram - a detailed laboratory study that evaluates the work of the gastrointestinal tract, the speed and efficiency of digestion and bowel function. Laboratory methods for examining feces reveal physical and Chemical properties feces, composition, presence of foreign organisms and inclusions. Requirements for stool collection:

• The material is taken after the natural act of defecation.
• The collection is carried out in a special container.
• It is forbidden to take biological material obtained as a result of an enema for examination of feces for dysentery.
• Before the study, it is forbidden to use iron preparations, put rectal suppositories, take laxatives and drink alcoholic beverages.

Microbiological diagnostics

Tank sowing for dysentery accurately determines the type of pathogen.

Bacteriological diagnostics - collection of fecal masses and subsequent sowing of feces in a special nutrient medium. The appearance of colonies of pathogenic bacteria (shigella) after sowing confirms the proposed diagnosis. Bacteriological analysis for dysentery accurately determines the pathogen, its type, subspecies and susceptibility to antibacterial agents, which allows you to choose the right drug for treatment.

The investigated material - feces with foreign impurities obtained naturally or a special tube for sigmoidoscopy. In children, a swab is taken with a special swab (a swab for the VD or a swab for the intestinal group). Establish sensitivity to drugs by placing colonies of Shigella along with various antibiotics. If the vital activity of microorganisms continues near the antibiotic tablet, then the drug is not used for treatment, if the microorganisms die, treatment with such an antibiotic is prescribed.

Serological tests for dysentery

In case of negative or doubtful results of bacteriological examination, a serological method is used. IN feces the patient is detected by a bacterial antigen, and in the plasma - specific antibodies. To establish the antibody titer, you can use the RIGA method, sometimes - RPGA or RA. A suspension of a daily colony of shegella is used as antigens. Minus the method - reliable results receive only 5 days after the onset of the disease, when the concentration of antibodies reaches the desired level.

Sigmoidoscopy

Due to the fact that the causative agent of dysentery affects the large intestine, sigmoidoscopy is a significant diagnostic method, but not a determining one. Diagnosis consists in introducing a rectoscope equipped with an air supply device into the anus. Swelling, the intestinal cavity becomes available for research. This method helps to assess the degree of damage to the intestinal epithelium. With dysentery, the intestinal walls are hyperemic as a result of vasodilation. Erosions and hemorrhages are formed on some segments. Sigmoidoscopy does not require preparation, but the procedure is not performed if there are anal fissures or pathologies of the anus.

Methodological instructions for students for practical lesson No. 28.

Lesson topic:

Target: Study of methods of microbiological diagnostics, etiotropic therapy and prevention of shigellosis.

Module 2 . Special, clinical and ecological microbiology.

Topic 5: Methods of microbiological diagnosis of dysentery.

Relevance of the topic:Shigellosis is ubiquitous and is serious problem in countries with a low sanitary cultural level and a high incidence of malnutrition and poor nutrition. In developing countries, the spread of infection is favored by poor sanitation, poor personal hygiene, overcrowding, and a large proportion of children in the population. In Ukraine, outbreaks of shigellosis are more common in closed communities with poor sanitation and hygiene, such as nurseries and kindergartens, tourist boats, psychiatric clinics or shelters for the disabled. Shigella has been the cause of travelers' and tourists' diarrhea.

The cause of group diseases can be considered the use of food products contaminated by the negligence of trade workers who are carriers of shigella. There are outbreaks associated with the use of drinking water, and swimming in polluted reservoirs also led to infection. However, the food and water routes of transmission seem to play a smaller role in the spread of shigellosis compared to cholera and typhoid fever, in which large doses of pathogens are usually required to infect a person. In developing countries, where the spread of the disease is predominantly person-to-person, carriers can be an important reservoir of the infectious agent. In patients who have not taken antibacterial drugs, shedding of shigella in the faeces usually lasts for 14 weeks, but in a small proportion of cases it lasts much longer.

Shigellosis is an acute bacterial infection of the intestine caused by one of the four types of Shigella. The spectrum of clinical forms of infection ranges from mild, watery diarrhea to severe dysentery characterized by abdominal cramps, tenesmus, fever, and signs of general intoxication.

Etiology.

The genus Shigella (named after K. Shiga, who in 1898 studied in detail and described the isolated causative agent of bacterial dysentery by A.V. Grigoriev) of the Enterobacteriaceae family consists of a group of closely related bacterial species with the following properties:

I. Morphological: shigella - small sticks with rounded ends. They differ from other representatives of the Enterobacteriaceae family in the absence of flagella (non-motile), do not have spores and capsules, and are gram-negative.

II. Cultural: shigella are aerobes or facultative anaerobes; optimal conditions cultivation temperature 37°C, pH 7.2-7.4. They grow on simple nutrient media (MPA, MPB) in the form of small, shiny, translucent, grayish, round colonies, 1.52 mm in size. S form. The exception is Sonne's Shigella, which often dissociate, forming large, flat, cloudy, jagged-edged colonies. R forms (colonies look like a “grape leaf”). In liquid nutrient media, Shigella give a uniform turbidity, R forms form a precipitate. The enrichment liquid medium is selenite broth.

III. Enzymatic: the main biochemical features necessary for the identification of shigella in the isolation of a pure culture are the following:

1. lack of gas formation during glucose fermentation;
2. no production of hydrogen sulfide;
3. no lactose fermentation within 48 hours.

Altogether, the four species are further subdivided into approximately 40 serotypes. According to the characteristics of the main somatic (O) antigens and biochemical properties, the following four species or groups are distinguished: S. dysenteriae (group A, includes: Grigoriev-Shigi, Stutzer-Schmitz, Large-Sachs), S. flexneri (group B), S. boydii (Group C) and S. sonnei (Group D).

In relation to mannitol, all shigella are divided into splitting (Flexner, Boyd, Sonne shigella) and non-splitting (Grigoriev-Shiga, Stutzer-Schmitz, Large-Sachs shigella) mannitol.

IV. Pathogenic factors:

1. Plasmid invasionprovides the ability of shigella to cause invasion with subsequent intercellular spread and reproduction in the epithelium of the colon mucosa;
2. toxin formation: Shigella have lipopolysaccharide endotoxin, which is chemically and biochemically similar to endotoxins of other members of the Enterobacteriaceae family. In addition, S. dysenteriae type I (Shiga's bacillus) produces an exotoxin. Since the discovery of the latter, it has been established that it has enterotoxin activity and can cause intestinal secretion, as well as have a cytotoxic effect against intestinal epithelial cells; has a neurotoxic effect, which is noted in children with shigellosis. Shiga toxin, getting into the blood, along with damage to the submucosal endothelium, also affects the glomeruli of the kidney, as a result of which, in addition to bloody diarrhea, hemolytic uremic syndrome develops with the development of renal failure.

V. Antigenic structure:All Shigella have a somatic O-antigen, depending on the structure of which they are divided into serovars.

VI. Resistance: Temperature 100 0 C kills shigella instantly. Shigella are resistant to low temperatures in river water they are stored up to 3 months, on vegetables and fruits - up to 15 months.Under favorable conditions, Shigella are able to reproduce in food products(salads, vinaigrettes, boiled meat, minced meat, boiled fish, milk and dairy products, compotes and jelly), especially Shigella Sonne.

Epidemiology.

1. Source of infection:A person suffering from acute and chronic forms of shigellosis; bacteriocarrier.

2. Ways of transmission:

• Food (mainly for S. sonnei)
• Aquatic (mainly for S. flexneri)
• Contact household (mainly for S. dysenteriae)

3. entrance gate infection serves the gastrointestinal tract.

Pathogenesis and pathological changes.

After ingestion, Shigella colonize the upper regions small intestine and multiply there, possibly causing increased secretion on early stage infections. Shigella then penetrate through the M cells into the submucosa, where they are engulfed by macrophages. This leads to the death of some of the shigella, resulting in the release of inflammatory mediators, which initiate inflammation in the submucosa. Apoptosis of phagocytes allows another part of the Shigella to survive and penetrate into the epithelial cells of the mucosa through the basement membrane. Inside the enterocytes, shigella reproduce and intercellular spread, resulting in the development of erosions. When shigella dies, shiga and shiga-like toxins are released, the action of which leads to intoxication. The defeat of the mucous membrane is accompanied by swelling, necrosis and hemorrhage, which causes the appearance of blood in the stool. In addition, the toxin affects the central nervous system, which leads to trophic disorders.

Clinical manifestations.

The spectrum of clinical manifestations of shigellosis is very wide from mild diarrhea to severe dysentery with cramping abdominal pain, tenesmus, fever and general intoxication.

Incubation periodranges from several hours to 7 days, most often it is 2-3 days.Initially, patients have watery stool, fever (up to 41 ° C), diffuse pain in the abdomen, nausea and vomiting. Along with this, patients complain of myalgia, chills, back pain and headache. In the coming days from the onset of the disease, signs of dysentery appear - tenesmus, frequent, scanty, bloody-mucous stools. Body temperature gradually decreases, pain can be localized in lower quadrants belly. The intensity of diarrhea reaches a maximum around the end of the 1st week of illness. Dysentery with bloody stools is more common and appears earlier in the disease caused by S. dysenteriae type I than in other forms of shigellosis.

For Shigellosis Sonne a milder course of the disease is characteristic (gastroenteric or gastroenterocolitic variant). The feverish period is shorter, the effects of intoxication are short-lived, and destructive changes in the intestinal mucosa are not typical.

Shigellosis FlexnerBasically, two variants of the clinical course are characteristic - gastroenterocolitic and colitis.

Extraintestinal complications in shigellosisrare:

1. A complication of shigellosis can be the development of intestinal dysbacteriosis.
2. Along with headaches, there may be signs of meningitis and convulsive seizures.
3. Cases of peripheral neuropathy have been described in S. dysenteriae type I infection, and cases of Guillain-Barré syndrome (polyneuritis) have been reported during an outbreak of S. boydii gastroenteritis.
4. With the exception of children suffering from dystrophy, hematogenous dissemination of the pathogen is relatively rare, and cases of shigellosis abscesses and meningitis have also been described.
5. With shigellosis, the development of Reiter's syndrome with arthritis, sterile conjunctivitis and urethritis is possible, this usually occurs after 1-4 weeks from the onset of diarrhea in patients.
6. In children, shigellosis is accompanied by a hemolytic uremic syndrome, often associated with leukemia-like reactions, severe colitis, and circulating endotoxin, but bacteremia is usually not detected.
7. Very rarely, purulent keratoconjunctivitis is caused by shigella that has entered the eyes as a result of self-infection with contaminated fingers.
8. Hypovolemic shock and DIC.
9. Peritonitis, intestinal gangrene, intestinal bleeding.

Immunity: Humans have a natural resistance to shigellosis. After past illness immunity is not stable, and after shigellosis Sonne is practically absent. With a disease caused by Shigella Grigoriev Shigi, a more stable antitoxic immunity is developed. The main role in protection against infection belongs to the secretory IgA , preventing adhesion, and cytotoxic antibody-dependent activity of intraepithelial lymphocytes, which, together with secretory IgA kill shigella.

Diagnostics and laboratory researches.

Purpose of the study: detection and identification of shigella for diagnosis; detection of bacteria carriers; detection of shigella in foods.

Research material: excrement, sectional material, foodstuffs.

Diagnostic methods:microbiological (bacteriological, microscopic (luminescent); serological; biological; allergy test.

Research progress:

1 day of study:Cultures should be done from freshly excreted feces or using rectal swabs (rectal tube); in the absence of suitable conditions, the material must be placed in a transport environment. For this, enteric agar (MacConkey or Shigella-Salmonella medium), moderately selective xylose-lysine-deoxycholate agar, KLD) and nutrient broth (selenite broth) should be used. If the time between collection and inoculation exceeds 2 hours, then preservative solutions should be used: 20% bile broth, combined Kauffmann medium.

• Excrements in the glycerin mixture are emulsified, a drop of the emulsion is applied to the medium and rubbed with a spatula. Differential media for Shigella are Ploskirev, Endo and EMS media (eosinmethylene blue agar). Ploskirev's medium (the composition of the medium includes: MPA, lactose, bile salts and the indicator brilliant green) is also an elective medium for shigella, because. inhibits the growth of Escherichia coli.
• In parallel with direct sowing, the collected material is sown on the enrichment medium - selenite broth.
• All crops are placed in a thermostat.

Day 2 of the study:

• The cups are removed from the thermostat, suspicious colonies are screened on Ressel's medium (nutrient medium which includes: agar-agar, Andrede indicator, 1% lactose, 0.1% glucose) and mannitol. Sowing is done by strokes on a sloping surface and an injection into an agar column. The inoculated Ressel medium is placed in a thermostat for 18-24 hours (in parallel, reseeding from the selenite medium to differential diagnostic media is done).
• Make smears (Gram stain), microscope.
• Prepare preparations "hanging" or "crushed" drop.
• Statement of indicative RA with polyvalent diagnostic shigellosis sera.
• Sowing suspicious colonies on agar slant.

Day 3 of the study:

• Microscopy of agar slant material.
• Cultures that did not ferment lactose on Ressel's medium are subjected to further study: smears are made (Gram stain), culture purity is checked. In the presence of gram-negative rods, inoculation is carried out on Hiss media, broth with indicator papers (to detect indole and hydrogen sulfide) and litmus milk.
• The inoculated media are placed in a thermostat for 18-24 hours.

Day 4 of the study:

• Accounting for a short "variegated series".
• Cultures suspicious for their enzymatic and cultural properties against Shigella are subjected to serological identification. Statement of RA on glass (typical and group diagnostic sera). Setting up the deployed RA.

As accelerated methods for shigellosis, applyfluorescence microscopy And biological sample(introduction of virulent strains of Shigella into the conjunctival sac (under the lower eyelid) of guinea pigs conjunctivitis develops by the end of the 1st day).

Allergic test Zuverkalovintradermal allergic test with dysentery (introduction of 0.1 ml of dysentery in the forearm positive reaction in case of infiltration and hyperemia). Allergological diagnostics is currently practically not used. Tsurvekalov's test does not differ in specificity, positive reactions are recorded not only in shigellosis, but also in salmonellosis, escherichiosis, yersiniosis, and other acute intestinal infections, and sometimes in healthy individuals.

Treatment and prevention.Bacteriophage is used for treatment and prevention according to epidemiological indications. oral administration, antibiotics after determining the antibiogram; in case of dysbacteriosis preparations of probiotics for the correction of microflora. To replenish the loss of fluids and electrolytes - the introduction of a glucose-electrolyte solution inside.

Specific goals:

Interpret the biological properties of pathogens of shigellosis.

Familiarize yourself with the classification of Shigella.

Learn to interpret the pathogenetic patterns of the infectious process caused by shigella.

To determine the methods of microbiological diagnostics, etiotropic therapy and prevention of shigellosis.

Be able to:

• Inoculate the test material on nutrient media.
  • Prepare smears and Gram stain.
  • Conduct microscopy of preparations using an immersion microscope.
  • Analyze morphological, cultural, enzymatic features of Shigella.

Theoretical questions:

1. Characteristics of pathogens of shigellosis. biological properties.

2. Classification of shigella. The underlying principles.

3. Epidemiology, pathogenesis and clinical features shigellosis.

4. Laboratory diagnostics.

5. Principles of treatment and prevention of shigellosis.

Practical tasks that are performed in class:

1. Microscopy of demonstration preparations from pure cultures of shigellosis pathogens.

2. Work on the bacteriological diagnosis of shigellosis: the study of fecal cultures on Ploskirev's medium.

3. Subculture of suspicious colonies on Ressel's medium and on BCH to determine indole formation and H 2 S .

4. Sketching of demonstration preparations and the scheme of microbiological diagnostics of shigellosis in the lesson protocol.

5. Registration of the protocol.

Literature:

1. Korotyaev A.I., Babichev S.A., Medical microbiology, immunology and virology / Textbook for medical universities, St. Petersburg "Special Literature", 1998. - 592p.

2. Timakov V.D., Levashev V.S., Borisov L.B. Microbiology / Textbook.-2nd ed., revised. And additional - M .: Medicine, 1983, -512s.

3. Pyatkin K.D. Krivoshein Yu.S. Microbiology with virology and immunology.- Kyiv: In and shcha school, 1992. - 431s.

4. Medical microbiology / Edited by V.I. Pokrovsky.-M.: GEOTAR-MED, 2001.-768s.

5. Guide to practical training in microbiology, immunology and virology. Ed. M.P. Zykov. M. "Medicine". 1977. 288 p.

6. Cherkes F.K., Bogoyavlenskaya L.B., Belskan N.A. Microbiology. / Ed. F.K. Circassian. M.: Medicine, 1986. 512 p.

7. Lecture notes.

Additional literature:

1. Makiyarov K.A. Microbiology, virology and immunology. Alma-Ata, "Kazakhstan", 1974. 372 p.

2. Titov M.V. Infectious diseases. - K., 1995. 321s.

3. Shuvalova E.P. infectious diseases. - M.: Medicine, 1990. - 559 p.

4. BME, Vol. 1, 2, 7.

5. Pavlovich S.A. Medical microbiology in graphs: Proc. allowance for medical in-comrade. Mn.: Vysh. school, 1986. 255 p.

Brief guidelines to work in the practical session.

At the beginning of the lesson, the level of preparation of students for the lesson is checked.

Independent work consists of studying the classification of shigella, analyzing the scheme of pathogenetic and clinical signs of shigellosis. Study of methods of laboratory diagnostics of shigellosis. Students carry out sowing of biomaterial on nutrient media. Then micropreparations are prepared, they are stained according to Gram, microscopy is performed, micropreparations are sketched and the necessary explanations are given. The composition of independent work also includes microscopy of demonstration preparations and their sketching in the protocol of the lesson.

At the end of the lesson, a test control and analysis of the final results of each student's independent work is carried out.

Technological map of the practical lesson.

p/n	Stages	Time in minutes	Ways of learning	Equipment	Location
	Checking and correcting the initial level of preparation for the lesson		Test tasks of the initial level	Tables, atlas	study room
	Independent work		Logical Structure Graph	Immersion microscope, dyes, glass slides, bacteriological loops, nutrient media, Ploskirev's medium, Ressel's medium, "variegated Hiss series"
	Self check and correction of mastering the material		Targeted learning tasks
	Test control		Tests
	Analysis of work results

Targeted learning tasks:

1. Feces were obtained from a child with acute intestinal infections (the collection of feces was carried out with a rectal tube) containing mucus and pus. What express diagnostic method should be used?

A. ELISA.

b. REEF.

C. RA.

D. RSK.

E. RIA.

1. The causative agent of dysentery was isolated from a sick child with acute intestinal infection. What morphological features are characteristic of the pathogen?

A . Gram-negative non-motile rod.

B . Gram-positive movable rod.

C . Forms a capsule on a nutrient medium.

D . Forms spores in the external environment.

E . Gram-positive streptobacilli.

3. A patient who fell ill three days ago and complains of a temperature of 38 ° C, abdominal pain, frequent liquid stool, the presence of blood in the stool, the doctor clinically diagnosed bacterial dysentery. What method of microbiological diagnostics should be used in this case and what material should be taken from the patient to confirm the diagnosis?

A. Bacterioscopic cal.

B. Bacteriological cal.

C. Bacterioscopic blood.

D. Bacteriological urine.

E. Serological blood.

4. Shigella Sonne was isolated from the patient's feces. What additional research is needed to establish the source of infection?

A . Carry out phage typing of the isolated pure culture.

B . Determine antibiogram.

C . Set up the precipitation reaction.

D . Set up the complement fixation reaction.

E . Set up a neutralization reaction.

5. Among the group of tourists (27 people) who used to drink water from the lake, after two days, 7 people developed symptoms of acute diarrhea. What material should be sent to the bacteriological laboratory to establish the etiology of this disease?

A. Water, feces of patients.

B. Water, the blood of the sick.

C. Food products.

D. Urine.

E. Phlegm.

6. A significant drawback of the microscopic diagnostic method for acute intestinal infections is its insufficient information content due to the morphological identity of bacteria of the family Enterobacteriaceae . What makes this method more informative?

A . Radioimmunoassay.

B . Coombs reaction.

C . Linked immunosorbent assay.

D . opsonization reaction.

E . Immunofluorescence reaction.

7. A 29-year-old patient was hospitalized with bouts of vomiting, diarrhea, and tenesmus. Feces with pieces of mucus and an admixture of blood. Bacteriological examination of bacteria from colonies on Ploskirev's medium revealed immobile, gram-negative rods that do not ferment lactose. Name the causative agent of the infectious process.

A. Shigella flexneri.

b. Vibrio eltor.

C. E. Coli.

D. Proteus mirabilis.

E. Salmonella enteritidis.

8. A lettuce was delivered to the microbiological laboratory, which is presumably the cause of an acute intestinal infection. What nutrient media is used for primary inoculation?

A . Yolk-salt agar, MPB.

b. MPA, MPB.

C . Selenite broth, Endo, Ploskireva.

D . Liver broth, Roux medium.

E . Blood agar, alkaline agar.

9. At microbiological research minced meat isolated bacteria belonging to the genus Shigella. The study of what properties of microbes led to such a conclusion?

A . Cultural, tinctorial.

B . Antigenic, cultural.

C . Saccharolytic, proteolytic.

D . Antigenic, immunogenic.

E . Morphological, antigenic.

10. Microscopic examination of vomit taken from a patient with symptoms of an acute intestinal infection revealed immobile rods. In what smear or preparation could bacterial motility be studied?

A . In a Gram-stained smear.

B . In a smear stained according to Tsil - Nelsen.

C . In the preparation "thick drop".

D . In a Neisser-stained smear.

E . In the preparation "crushed drop".

Algorithm laboratory work :

1. Study of the biological properties of Shigella.

2. Acquaintance with the classification of shigella.

3. Analysis of the scheme of pathogenetic and clinical manifestations of shigellosis.

4. Study of methods of laboratory diagnostics of shigellosis.

5. Studying the basic principles of therapy and prevention of shigellosis.

1. Preparation of fixed preparations from bacterial culture.
2. Coloring micropreparations by Gram.
3. Microscopy of micropreparations With using an immersion microscope, their analysis and sketching in the protocol of the lesson.
4. Mi chromoscopy and analysis of demonstration preparations from pure cultures of Shigella.
5. Sketching of demonstration preparations and scheme of laboratory diagnostics of shigellosis in the protocol.
6. Formulation of the protocol.

Classification of shigella, their properties. The pathogenesis of shigellosis.

Bacterial dysentery, or shigellosis, is an infectious disease caused by bacteria of the genus Shigella, occurring with a predominant lesion of the colon. The name of the genus is associated with K. Shigi, who discovered one of the pathogens

dysentery.

Taxonomy and classification. The causative agents of dysentery belong to the Gracilicutes department, the Enterobacteriaceae family, the Shigella genus.

Morphology and tinctorial properties. Shigella - gram-negative rods with rounded ends, 2-3 microns long, 0.5-7 microns thick (see Fig. 10.1); do not form spores, do not have flagella, are immobile. In many strains, villi of a general type and genital pili are found. Some Shigella have a microcapsule.

Cultivation. Dysentery sticks are facultative anaerobes. They are undemanding to nutrient media, grow well at a temperature of 37 ° C and a pH of 7.2-7.4. On dense media they form small transparent colonies, in liquid media -

diffuse haze. Selenite broth is most often used as an enrichment medium for the cultivation of Shigella.

Enzymatic activity. Shigella have less enzymatic activity than other enterobacteria. They ferment carbohydrates with the formation of acid. An important feature that makes it possible to differentiate Shigella is their relationship to mannitol: S. dysenteriae do not ferment mannitol, representatives of groups B, C, D are mannitol-positive. The most biochemically active are S. sonnei, which slowly (within 2 days) can ferment lactose. Based on the relationship of S. sonnei to rhamnose, xylose and maltose, 7 biochemical variants of it are distinguished.

Antigenic structure. Shigella have O-antigen, its heterogeneity allows serovars and subserovars to be distinguished within groups; in some members of the genus, the K-antigen is found.

pathogenicity factors. All dysenteric bacilli form endotoxin, which has an enterotropic, neurotropic, pyrogenic effect. In addition, S. dysenteriae (serovar I) - shigella Grigoriev-Shigi - secrete an exotoxin that has an enterotoxic, neurotoxic, cytotoxic and nephrotoxic effect on the body, which accordingly disrupts water-salt metabolism and the activity of the central nervous system, leads to the death of epithelial cells of the colon, damage to the renal tubules. More than severe course dysentery caused by this pathogen. Other types of Shigella can secrete exotoxin. The RF permeability factor has been discovered, as a result of which blood vessels are affected. Pathogenicity factors also include an invasive protein that facilitates their penetration into epithelial cells, as well as pili and outer membrane proteins responsible for adhesion, and a microcapsule.

resistance. Shigella have a low resistance to action various factors. S. sonnei are more resistant, which persist in tap water for up to 2 "/2 months, in the water of open reservoirs they survive up to V / 2 months. S. sonnei can not only persist for a long time, but also multiply in products, especially dairy products.

Epidemiology. Dysentery is an anthroponotic infection: the source is sick people and carriers. The mechanism of transmission of infections is fecal-oral. The routes of transmission can be different - with Sonne's dysentery, the food route predominates, with Flexner's dysentery - water, for Grigoriev-Shiga's dysentery, the contact-household route is characteristic. Dysentery occurs in many countries of the world. In recent

years, there has been a sharp rise in the incidence of this infection. People of all ages get sick, but children from 1 to 3 years old are most susceptible to dysentery. The number of patients increases in July - September. Different types of shigella by individual

regions are unevenly distributed.

Pathogenesis. Shigella enters the gastrointestinal tract through the mouth and reaches the large intestine. Possessing tropism for its epithelium, pathogens attach to cells with the help of pili and proteins of the outer membrane. Thanks to the invasive factor, they penetrate inside the cells, multiply there, as a result of which the cells die. Ulcerations form in the intestinal wall, in place of which scars are then formed. Endotoxin, released during the destruction of bacteria, causes general intoxication, increased intestinal motility, and diarrhea. Blood from the formed ulcers enters the stool. As a result of the action of exotoxin, a more pronounced violation of water-salt metabolism, the activity of the central nervous system, and kidney damage is observed.

clinical picture. The incubation period lasts from 1 to 5 days. The disease begins acutely with an increase in body temperature to 38-39 ° C, abdominal pain, diarrhea appear. An admixture of blood, mucus is found in the stool. The most severe dysentery is Grigoriev-Shiga.

Immunity. After a disease, immunity is not only species-specific, but also variant-specific. It is short lived and unstable. Often the disease becomes chronic.

Microbiological diagnostics. The stool of the patient is taken as the test material. The basis of diagnosis is the bacteriological method, which allows to identify the pathogen, to determine its sensitivity to

antibiotics, conduct intraspecific identification (determine the biochemical variant, serovar or colicinogenovar). With a protracted course of dysentery, it can be used as an auxiliary serological method, which consists in staging RA, RNHA (by increasing the antibody titer during repeated reaction, the diagnosis can be confirmed).

Treatment. Patients with severe forms of dysentery Grigoriev-Shiga and Flexner are treated with broad-spectrum antibiotics with the obligatory consideration of the antibiogram, since among Shigella there are often not only antibiotic-resistant

chivy, but also antibiotic-dependent forms. In mild forms of dysentery, antibiotics are not used, since their use leads to dysbacteriosis, which makes it harder pathological process, and disruption of recovery processes in the mucous membrane of the colon.

Prevention. The only drug that can be used in the foci of infection for prophylactic purposes is dysenteric bacteriophage. The main role is played by non-specific prophylaxis.

11. Yersinia - the causative agents of the plague. Properties. Pathogenesis, immunity, laboratory diagnostics, epidemiology, prevention, treatment. The role of domestic scientists in the study of plague.

Taxonomy: Y.pestis causes plague; department Gracilicutes, family Enterobacteriaceae, genus Yersinia. The causative agent is Yersinia pestis.

Morphological properties: Gram-negative rods, ovoid, stain bipolar. They are mobile, have a capsule, do not form spores.

cultural properties.

facultative anaerobes. Temperature optimum + 25С. Well cultivated on simple nutrient media. Most carbohydrates are fermented without the formation of gas. Psychophiles - able to change their metabolism depending on temperature and multiply at low temperatures. Virulent strains form rough (R) colonies, transitional (RS) and grayish slimy smooth avirulent (S) forms.

Two types of colonies - young and mature. Juvenile with irregular margins. Mature colonies are large, with a brown granular center and jagged edges. On slant agar, a turn of two days at +28 C form a grayish - white coating, growing into the environment, on the broth - a delicate surface film and a cottony precipitate.

Biochemical properties: high enzyme activity: fermentation to acid xylose, synthesis of plasmacoagulase, fibrinolysin, hemolysin, lecithinase, hydrogen sulfide. Rhamnose, urea does not ferment.

Antigenic structure.

A group of protein - polysaccharide and lipopolysaccharide antigens: thermostable somatic O-antigen and thermolabile capsular V, W antigens. The virulence of bacteria is associated with the W antigen. It produces pathogenicity factors: fibrinolysin, plasmacoagulase, endotoxin, exotoxin, capsule, V, W antigens.

Resistance: sensitive to antibiotics (especially streptomycin), unstable to the environment at high temperatures.

pathogenic properties.

Possesses pathogenic potential, suppresses functions phagocytic system, suppresses the oxidative burst in phagocytes and multiplies freely in them. Pathogenicity factors are controlled by three classes of plasmids. In the pathogenesis, there are three main stages - lymphogenous drift, bacteremia, generalized septicemia. They have adhesins and invasins, low molecular weight proteins (inhibit bactericidal factors), enterotoxin. Some factors are controlled by virulence plasmids.

Clinical features: The incubation period is several hours up to 8 days. Distinguish local - skin-bubonic, bubonic; externally disseminated - primary pulmonary, secondary pulmonary and intestinal; generalized - primary septic, secondary septic forms of plague. Regional lymphadenopathy, enterocolitis, reactive arthritis, spondylitis, fever.

Epidemiology: Plague is a classic natural focal zoonosis of wild animals. The main carriers in nature are marmots, ground squirrels, in urban conditions - rats. In the transmission of the pathogen - fleas of animals that can infect humans.

Immunity: cellular-humoral, limited in duration and intensity.

Microbiological diagnostics:

Bacterioscopic examination. Smears are prepared from the test material, stained by Gram and an aqueous solution of methylene blue. Plague bacteria are gram-negative, ovoid-shaped rods. bacteriological research. The test material was inoculated onto nutrient agar plates. The cultures are incubated at 25C. The primary study of crops is carried out after 10 hours. By this time, colonies appear that are formed by virulent R-forms. Low and avirulent bacteria form S-shaped colonies. Identification of a pure culture is carried out according to the morphology of bacterial cells, the nature of growth, antigenic and biochemical properties, sensitivity to a specific phage and bioassay.

Bacteria form a film on the broth; ferment many sugars to acid, do not form indole, do not liquefy gelatin. They contain a group thermostable somatic antigen and a specific thermolabile capsular antigen.

Bioassay. It is carried out to isolate a pure culture from a material contaminated with foreign microflora. The most sensitive laboratory animals are guinea pigs, to which the material is injected subcutaneously. Intraperitoneally, the material is injected if it is not contaminated with other bacteria. After the death of animals, pathological changes in organs are noted and bacteriological examination is carried out.

Express methods of laboratory diagnostics:

2.RPGA - for the detection of bacterial antigens in the material using standard anti-plague serum, the antibodies of which are loaded on erythrocytes.

Treatment: antibiotics - streptomycin, tetracycline drugs.

Prevention: specific prophylaxis - live attenuated EV plague vaccine. A dry tablet vaccine is available for oral administration. To assess immunity to plague (natural post-infection and vaccine), an intradermal allergy test with pestin can be used.

Plague bacteriophage– when identifying Y.pestis.

Plague dry vaccine - dried live culture of Y. pestis vaccine strain EV, used for the prevention of plague.

Microbiology of dysentery

Dysentery is an infectious disease characterized by general intoxication of the body, diarrhea and a peculiar lesion of the mucous membrane of the large intestine. It is one of the most frequent acute intestinal diseases in the world. The disease has been known since ancient times under the name of "bloody diarrhea", but its nature turned out to be different. In 1875, the Russian scientist F. A. Lesh isolated an amoeba from a patient with bloody diarrhea Entamoeba histolytica, in the next 15 years, the independence of this disease was established, for which the name amoebiasis was preserved.

The causative agents of dysentery proper are a large group of biologically similar bacteria united in the genus Shigella. The pathogen was first discovered in 1888 by A. Chantemes and F. Vidal; in 1891, it was described by A. V. Grigoriev, and in 1898, K. Shiga, using the serum he received from a patient, identified the pathogen in 34 patients with dysentery, finally proving the etiological role of this bacterium. However, in subsequent years, other causative agents of dysentery were also discovered: in 1900 - by S. Flexner, in 1915 - by K. Sonne, in 1917 - by K. Stutzer and K. Schmitz, in 1932 - by J. Boyd , in 1934 - by D. Large, in 1943 - by A. Sachs. Currently the genus Shigella includes more than 40 serotypes. They are all short immobile gram-negative rods that do not form spores and capsules, which grow well on ordinary nutrient media, do not grow on a starvation medium with citrate or malonate as the sole carbon source; do not form H 2 S, do not have urease; the Voges-Proskauer reaction is negative; glucose and some other carbohydrates are fermented to form acid without gas (except for some biotypes Shigella flexneri: S. manchester And S. newcastle); as a rule, do not ferment lactose (with the exception of Shigella Sonne), adonite, salicin and inositol, do not liquefy gelatin, usually form catalase, do not have lysine decarboxylase and phenylalanine deaminase. The content of G + C in DNA is 49 - 53 mol%. Shigella are facultative anaerobes, the temperature optimum for growth is 37 °C, they do not grow at temperatures above 45 °C, the optimum pH of the medium is 6.7 - 7.2. Colonies on dense media are round, convex, translucent; in the case of dissociation, rough R-shaped colonies are formed. Growth on the BCH in the form of a uniform turbidity, rough forms form a precipitate. Freshly isolated cultures of Sonne Shigella usually form colonies of two types: small round convex (I phase), large flat (II phase). The nature of the colony depends on the presence (I phase) or absence (II phase) of a plasmid with m. m. 120 MD, which also determines the virulence of Shigella Sonne.

International classification Shigella is built taking into account their biochemical characteristics (mannitol-non-fermenting, mannitol-fermenting, slowly lactose-fermenting Shigella) and features of the antigenic structure (Table 37).

O-antigens of different specificity were found in Shigella: common for the family Enterobacteriaceae, generic, species, group and type-specific, as well as K-antigens; They do not have H antigens.

Table 37

Classification of bacteria of the genus Shigella

The classification takes into account only group and type-specific O-antigens. According to these features, the Shigella subdivided into 4 subgroups, or 4 species, and includes 44 serotypes. In subgroup A (species Shigella dysenteriae) includes shigella that do not ferment mannitol. The species includes 12 serotypes (1 - 12). Each serotype has its own specific type antigen; antigenic relationships between serotypes, as well as with other types of shigella, are weakly expressed. To subgroup B (type Shigella flexneri) include Shigella, usually fermenting mannitol. Shigella of this species are serologically related to each other: they contain type-specific antigens (I - VI), according to which they are divided into serotypes (1 - 6), and group antigens, which are found in different compositions in each serotype and according to which serotypes are divided into subserotypes. In addition, this species includes two antigenic variants, X and Y, which do not have typical antigens; they differ in sets of group antigens. Serotype S. flexneri 6 does not have subserotypes, but it is divided into 3 biochemical types according to the characteristics of the fermentation of glucose, mannitol and dulcite (Table 38).

Table 38

Biotypes S. flexneri 6

Note. K - fermentation with the formation of only acid; KG - fermentation with the formation of acid and gas; (-) - no fermentation.

The lipopolysaccharide antigen O in all Shigella Flexner contains group antigen 3, 4 as the main primary structure, its synthesis is controlled by a chromosomal gene localized near the his-locus. Type-specific antigens I, II, IV, V and group antigens 6, 7, 8 are the result of modification of antigens 3, 4 (glycosylation or acetylation) and are determined by the genes of the corresponding converting prophages, the integration site of which is located in the lac-pro region of the Shigella chromosome.

Appeared on the territory of the country in the 80s. 20th century and a widely used new subserotype S. flexneri 4(IV:7, 8) differs from subserotype 4a (IV:3, 4) and 4b (IV:3, 4, 6), arose from the variant S. flexneri Y(IV:3, 4) due to lysogenization by its converting prophages IV and 7, 8.

To subgroup C (kind Shigella boydii) include Shigella, usually fermenting mannitol. Members of the group are serologically distinct from each other. Antigenic relationships within the species are weakly expressed. The species includes 18 serotypes (1 - 18), each of which has its own main type antigen.

In subgroup D (species Shigella sonnei) includes Shigella, which usually ferment mannitol and are able to slowly (after 24 h of incubation and later) ferment lactose and sucrose. View S. sonnei includes one serotype, however, phases I and II colonies have their own type-specific antigens. Two methods have been proposed for the intraspecific classification of Sonne's Shigella:

1) dividing them into 14 biochemical types and subtypes according to their ability to ferment maltose, rhamnose and xylose; 2) division into phage types according to sensitivity to a set of corresponding phages.

These typing methods are mainly of epidemiological significance. In addition, Sonne's shigella and Flexner's shigella are subjected to typing for the same purpose by the ability to synthesize specific colicins (colicinogenotyping) and by sensitivity to known colicins (colicinotyping). To determine the type of colicins produced by shigella, J. Abbott and R. Shannon proposed sets of typical and indicator strains of shigella, and to determine the sensitivity of shigella to known types of colicins, a set of reference colicinogenic strains by P. Frederick is used.

resistance. Shigella have a fairly high resistance to environmental factors. They survive on cotton fabric and on paper up to 30-36 days, in dried feces - up to 4-5 months, in soil - up to 3-4 months, in water - from 0.5 to 3 months, on fruits and vegetables - up to 2 weeks, in milk and dairy products - up to several weeks; at a temperature of 60 ° C they die in 15 - 20 minutes. Sensitive to chloramine solutions, active chlorine and other disinfectants.

pathogenicity factors. The most important biological property of Shigella, which determines their pathogenicity, is the ability to invade epithelial cells, multiply in them and cause their death. This effect can be detected using a keratoconjunctival test (the introduction of one loop of a Shigella culture (2–3 billion bacteria) under the lower eyelid of a guinea pig causes the development of serous purulent keratoconjunctivitis), as well as by infection of cell cultures (cytotoxic effect) or chicken embryos (their death) , or intranasally in white mice (development of pneumonia). The main pathogenicity factors of shigella can be divided into three groups:

1) factors that determine the interaction with the epithelium of the mucous membrane;

2) factors that provide resistance to humoral and cellular defense mechanisms of the macroorganism and the ability of Shigella to multiply in its cells;

3) the ability to produce toxins and toxic products that determine the development of the actual pathological process.

The first group includes adhesion and colonization factors: their role is played by pili, outer membrane proteins, and LPS. Mucus-destroying enzymes such as neuraminidase, hyaluronidase, and mucinase promote adhesion and colonization. The second group includes invasion factors that promote the penetration of Shigella into enterocytes and their reproduction in them and in macrophages with the simultaneous manifestation of a cytotoxic and (or) enterotoxic effect. These properties are controlled by the genes of the plasmid with m. m. 140 MD (it encodes the synthesis of proteins of the outer membrane that cause invasion) and the Shigella chromosomal genes: kcp A (causes keratoconjunctivitis), cyt (responsible for cell destruction), as well as other genes, not yet identified. Protection of Shigella from phagocytosis is provided by surface K-antigen, antigens 3, 4 and lipopolysaccharide. In addition, Shigella endotoxin lipid A has an immunosuppressive effect: it suppresses the activity of immune memory cells.

The third group of pathogenicity factors includes endotoxin and two types of exotoxins found in Shigella - Shiga exotoxins and Shiga-like exotoxins (SLT-I and SLT-II), whose cytotoxic properties are most pronounced in S. dysenteriae 1. Shiga- and Shiga-like toxins also found in other serotypes S. dysenteriae, they are also formed S. flexneri, S. sonnei, S. boydii, EHEC and some salmonella. The synthesis of these toxins is controlled by the tox genes of converting phages. Type LT enterotoxins have been found in Flexner, Sonne and Boyd Shigella. Synthesis of LT in them is controlled by plasmid genes. Enterotoxin stimulates the activity of adenylate cyclase and is responsible for the development of diarrhea. Shiga toxin, or neurotoxin, does not react with the adenylate cyclase system, but has a direct cytotoxic effect. Shiga and Shiga-like toxins (SLT-I and SLT-II) have a MW of 70 kD and consist of A and B subunits (the last of 5 identical small subunits). The receptor for toxins is the glycolipid of the cell membrane.

The virulence of Shigella Sonne also depends on the plasmid with m. m. 120 MD. It controls the synthesis of about 40 outer membrane polypeptides, seven of which are associated with virulence. Shigella Sonne with this plasmid form phase I colonies and are virulent. Cultures that have lost the plasmid form phase II colonies and lack virulence. Plasmids with m.m. 120 - 140 MD were found in Shigella Flexner and Boyd. Shigella lipopolysaccharide is a potent endotoxin.

Features of epidemiology. The only source of infection is humans. No animal in nature suffers from dysentery. Under experimental conditions, dysentery can only be reproduced in monkeys. The method of infection is fecal-oral. Ways of transmission - water (predominant for Shigella Flexner), food, especially important role belongs to milk and dairy products (the predominant route of infection for Shigella Sonne), and contact-household, especially for the species S. dysenteriae.

A feature of the epidemiology of dysentery is the change in the species composition of pathogens, as well as Sonne biotypes and Flexner serotypes in certain regions. For example, until the end of the 1930s 20th century to share S. dysenteriae 1 accounted for up to 30 - 40% of all cases of dysentery, and then this serotype began to occur less and less and almost disappeared. However, in the 1960s - 1980s. S. dysenteriae reappeared on the historical arena and caused a series of epidemics that led to the formation of three hyperendemic foci of it - in Central America, Central Africa and South Asia (India, Pakistan, Bangladesh and other countries). The reasons for the change in the species composition of dysentery pathogens are probably associated with a change in collective immunity and with a change in the properties of dysentery bacteria. In particular, the return S. dysenteriae 1 and its wide distribution, which caused the formation of hyperendemic foci of dysentery, is associated with the acquisition of plasmids by it, which caused multiple drug resistance and increased virulence.

Features of pathogenesis and clinic. The incubation period for dysentery is 2-5 days, sometimes less than a day. Formation infectious focus in the mucous membrane of the descending part of the large intestine (sigmoid and rectum), where the causative agent of dysentery penetrates, is cyclic: adhesion, colonization, the introduction of shigella into the cytoplasm of enterocytes, their intracellular reproduction, destruction and rejection of epithelial cells, the release of pathogens into the intestinal lumen; after this, the next cycle begins - adhesion, colonization, etc. The intensity of the cycles depends on the concentration of pathogens in the parietal layer of the mucous membrane. As a result of repeated cycles, the inflammatory focus grows, the resulting ulcers, connecting, increase exposure intestinal wall, as a result of which blood, mucopurulent lumps, polymorphonuclear leukocytes appear in the stool. Cytotoxins (SLT-I and SLT-II) cause cell destruction, enterotoxin - diarrhea, endotoxins - general intoxication. The clinic of dysentery is largely determined by what type of exotoxins is produced to a greater extent by the pathogen, the degree of its allergenic effect and the body's immune status. However, many issues of the pathogenesis of dysentery remain unexplained, in particular: the course of dysentery in children of the first two years of life, the reasons for the transition of acute dysentery to chronic, the significance of sensitization, the mechanism of local immunity of the intestinal mucosa, etc. The most typical clinical manifestations of dysentery are diarrhea, frequent urges: in severe cases up to 50 or more times a day, tenesmus (painful spasms of the rectum) and general intoxication. The nature of the stool is determined by the degree of damage to the large intestine. The most severe dysentery is caused by S. dysenteriae 1, most easily - Sonne's dysentery.

Postinfectious immunity. As observations on monkeys have shown, after suffering dysentery, a strong and fairly long-term immunity remains. It is caused by antimicrobial antibodies, antitoxins, increased activity of macrophages and T-lymphocytes. A significant role is played by local immunity of the intestinal mucosa, mediated by IgAs. However, immunity is type-specific in nature, strong cross-immunity does not occur.

Laboratory diagnostics. The main method is bacteriological. The material for the study is feces. Pathogen isolation scheme: inoculation on the Endo and Ploskirev differential diagnostic media (in parallel on the enrichment medium, followed by inoculation on the Endo and Ploskirev media) to isolate isolated colonies, obtaining a pure culture, studying its biochemical properties and, taking into account the latter, identification using polyvalent and monovalent diagnostic agglutinating sera. The following commercial serums are produced.

1. Shigella that do not ferment mannitol:

To S. dysenteriae 1 And 2

To S. dysenteriae 3–7(polyvalent and monovalent),

To S. dysenteriae 8 – 12(polyvalent and monovalent).

2. To shigella fermenting mannitol:

to typical antigens S. flexneri I, II, III, IV, V, VI,

to group antigens S. flexneri 3, 4, 6, 7, 8- polyvalent,

to antigens S.boydii 1–18(polyvalent and monovalent), to antigens S. sonnei I phase, II phase,

to antigens S. flexneri I–VI+ S. sonnei- polyvalent.

For quick identification of Shigella, the following method is recommended: a suspicious colony (lactosonegative on Endo medium) is subcultured on TSI medium (eng. triple sugar iron) - three-sugar agar (glucose, lactose, sucrose) with iron to determine the production of H 2 S; or on a medium containing glucose, lactose, sucrose, iron and urea. Any organism that breaks down urea after 4 to 6 hours of incubation is most likely to belong to the genus Proteus and may be excluded. A microorganism producing H 2 S or having urease or producing acid at the joint (ferments lactose or sucrose) can be excluded, although H 2 S producing strains should be investigated as possible members of the genus Salmonella. In all other cases, the culture grown on these media should be examined and, if it ferments glucose (color change of the bar), isolated in pure form. At the same time, it can be investigated in the agglutination test on glass with the appropriate antisera to the genus Shigella. If necessary, other biochemical tests are carried out to check belonging to the genus Shigella and study mobility.

The following methods can be used to detect antigens in the blood (including as part of the CEC), urine and feces: RPHA, RSK, coagglutination reaction (in urine and feces), IFM, RAGA (in blood serum). These methods are highly effective, specific and suitable for early diagnosis.

For serological diagnosis, the following can be used: RPGA with the corresponding erythrocyte diagnosticums, immunofluorescent method (in indirect modification), Coombs method (determination of the titer of incomplete antibodies). An allergic test with dysentery (a solution of protein fractions of Shigella Flexner and Sonne) is also of diagnostic value. The reaction is taken into account after 24 hours. It is considered positive in the presence of hyperemia and infiltration with a diameter of 10-20 mm.

Treatment. The main attention is paid to the restoration of normal water-salt metabolism, rational nutrition, detoxification, rational antibiotic therapy (taking into account the sensitivity of the pathogen to antibiotics). A good effect is obtained by the early use of a polyvalent dysenteric bacteriophage, especially tablets with a pectin coating, which protects the phage from the action of HCl of gastric juice; in the small intestine, pectin dissolves, phages are released and show their action. For prophylactic purposes, the phage should be given at least once every three days (the period of its survival in the intestine).

The problem of specific prevention. To create artificial immunity against dysentery, various vaccines were used: from killed bacteria, chemical, alcohol, but all of them turned out to be ineffective and were discontinued. Vaccines against Flexner's dysentery have been created from live (mutant, streptomycin-dependent) Shigella Flexner; ribosomal vaccines, but they have also not been widely used. Therefore, the problem of specific prevention of dysentery remains unresolved. The main way to combat dysentery is to improve the water supply and sewerage system, ensure strict sanitary and hygienic regimes in food enterprises, especially the dairy industry, in childcare facilities, public places and in personal hygiene.

UDC 616.935-074(047)

A.M.Sadykova

Kazakh National Medical University

named after S.D. Asfendiyarov, Almaty

Department of Infectious and Tropical Diseases

Reliable diagnosis of dysentery is one of the urgent tasks of AEI surveillance. An accurate diagnosis of bacillary dysentery is importance for the right and timely treatment patient and for the implementation of the necessary anti-epidemic measures. The data presented in the review show that, given the widespread prevalence of dysentery, lack of sensitivity and late appearance positive results of many diagnostic methods, it is advisable to develop the diagnostic potential of detecting this infection.

Keywords: diagnostics, dysentery, antigen-binding lymphocyte method.

Recognition of shigellosis infection in clinical practice encounters significant difficulties due to objective factors, which include the clinical pathomorphism of dysentery, an increase in the number atypical forms diseases, the existence of a significant number of diseases of an infectious and non-infectious nature, having clinical manifestations similar to dysentery. Under the diagnosis of "clinical dysentery" in half of the cases, unrecognized diseases of a different etiology are hidden.

The greatest difficulties arise before the doctor during the initial examination of the patient before obtaining the results of paraclinical diagnostic methods. Recognition of dysentery is also difficult in the presence of concomitant diseases gastrointestinal tract.

Since the beginning of the use of the etiological laboratory diagnosis of dysentery, quite a few methods have been proposed and tested. There are many classifications of methods for the etiological diagnosis of infections. Methodologically, the classification proposed by B.V. Punishment. With regard to the diagnosis of dysentery, the principles of methodologically sound classification were used by B.V. Karalnik, N.M. Nurkina, B.K. Erkinbekova..

Of the laboratory methods for diagnosing dysentery, bacteriological (isolation and identification of the pathogen) and immunological are known. The latter include immunological methods in vivo (allergological test Zuverkalov) and in vitro. Immunological methods in vitro have one undoubted advantage over the Zuverkalov test - they are not associated with the introduction of foreign antigens into the body.

Most researchers still believe that bacteriological research, which includes isolation in pure culture the causative agent of the disease with its subsequent identification by morphological, biochemical and antigenic characteristics, is the most reliable method for diagnosing shigellosis infection. The frequency of isolation of shigella from the feces of patients with a clinical diagnosis of "acute dysentery", according to different authors, ranges from 30.8% to 84.7% and even 91.1%. Such a significant range for different authors depends not only on objective factors affecting the effectiveness of bacteriological examination, but also on the thoroughness of the diagnosis (or exclusion) of "clinical dysentery". The effectiveness of bacteriological research is influenced by such objective factors, as features of the course of the disease, the method of sampling and delivery of material to the laboratory, the quality of nutrient media, the qualifications of personnel, the timing of the patient's contact with health workers, the use antimicrobials before taking material for research. quantitative microbiological research feces in acute dysentery shows that in any clinical forms of infection, the most massive release of pathogens occurs in the first days of the disease, and starting from the 6th and, especially from the 10th day of the disease, the concentration of shigella in the feces is significantly reduced. T.A. Avdeeva found that the low content of shigella and the sharp predominance of non-pathogenic microorganisms in the feces practically exclude the possibility of bacteriological detection of dysentery bacteria.

It is known that bacteriological confirmation of shigellosis infection is most often successful when examining patients in the first days of the disease - the coproculture of the pathogen in the vast majority of cases is first isolated during the first study. Positive results of bacteriological examination are noted only in the first 3 days of the disease in 45 - 49% of patients, in the first 7 days - in 75%. Tillet and Thomas also consider the timing of the examination of patients as an important factor in determining the effectiveness of the bacteriological method for diagnosing dysentery. According to T.A. Avdeeva, in the first days of the disease, the most intense release of the pathogen is observed with Sonne dysentery, less intense with Flexner dysentery and the least with Flexner VI dysentery; in the later stages of the disease, the highest concentration is maintained for the longest time in Flexner's dysentery, less long - Shigella Sonne and the least long - Shigella Flexner VI.

Thus, although the bacteriological examination of feces is the most reliable method for diagnosing shigellosis infection, the limitations of its effectiveness listed above are significant drawbacks. It is also important to point out the limitations of early diagnosis by the bacteriological method, in which the duration of the analysis is 3-4 days. In connection with these circumstances, the use of other methods of laboratory diagnostics is of great practical importance. Another microbiological method for diagnosing dysentery is also based on the detection of live Shigella. This is a phage titer rise reaction (RNF) based on the ability of specific phages to multiply exclusively in the presence of homologous live microorganisms. An increase in the indicator phage titer indicates the presence of the corresponding microbes in the medium. The diagnostic value of the RNF in shigellosis infection was tested by B.I. Khaimzon, T.S. Vilkomirskaya. RNF has a fairly high sensitivity. Mapping minimum concentration Shigella in stools captured by the bacteriological method (12.5 thousand bacteria in 1 ml) and RNF (3.0 - 6.2 thousand) indicates the superiority of RNF.

Since the frequency of positive RNF results is directly dependent on the degree of contamination of the feces, the use of the method also gives the greatest effect in the first days of the disease and in more severe forms of the infectious process. However, the higher sensitivity of the method causes its special advantages over bacteriological examination in the late stages of the disease, as well as in the examination of patients with mild, asymptomatic and subclinical forms of infection, with a low concentration of the pathogen in the feces. RNF is also used in the examination of patients taking antibacterial agents, since the latter drastically reduce the frequency of positive results of the bacteriological method of research, but to a much lesser extent affect the effectiveness of the RNF. The sensitivity of the RNF is not absolute due to the existence of phage-resistant strains of shigella: the proportion of phage-resistant strains can vary over a very wide range - from 1% to 34.5%.

The great advantage of RNF is its high specificity. When examining healthy people, as well as patients with infectious diseases of a different etiology, positive reaction results were observed only in 1.5% of cases. RNF is a valuable additional method for diagnosing shigellosis infection. But today this method is rarely used because of its technical complexity. Other methods are immunological. With their help, a specific immune response is registered with respect to the pathogen or the antigens of the pathogen are determined by immunological methods.

Due to the severity of the processes of specific infectious allergy in shigellosis infection, allergological diagnostic methods were first used, which include an intradermal allergic test with dysentery (VPD). The drug "dysentery", which is a specific Shigella allergen devoid of toxic substances, was obtained by D.A. Tsuverkalov and was first used in clinical conditions when setting up an intradermal test by L.K. Korovitsky in 1954. According to E.V. Golyusova and M.Z. Trokhimenko, in the presence of previous acute dysentery or associated allergic diseases with skin manifestations (eczema, urticaria, etc.). positive results of VPD are observed much more often (paraallergy). Analysis of the results of the VPD in different periods acute dysentery shows that a specific allergy occurs already in the first days of the disease, reaches its maximum severity by the 7th - 15th day and then gradually fades away. Positive reaction results were obtained when examining healthy people aged 16 to 60 years in 15 - 20% of cases and aged 3 to 7 years - in 12.5% ​​of cases. Even more often, non-specific positive results of VPD were observed in patients with gastrointestinal diseases - in 20 - 36% of cases. The introduction of the allergen was accompanied by the development of a local reaction in 35.5 - 43.0% of patients with salmonellosis, in 74 - 87% of patients with coli-0124-enterocolitis. A serious argument against the widespread use of VPD in clinical practice was its allergenic effect on the body. Given the above, we can say that this method is not very specific. Tsuverkalov's test is also not species-specific. Positive reaction results were equally frequent in various etiological forms of dysentery.

In addition to VPD, other diagnostic reactions were also used, with varying degrees of validity, considered as allergic, for example, the allergen leukocytolysis (ALC) reaction, the essence of which was the specific damage or complete destruction of actively or passively sensitized neutrophils upon contact with the corresponding AG. But this reaction cannot be attributed to the methods of early diagnosis, since the maximum frequency of positive results was observed on the 6-9th day of the disease and amounted to 69%. An allergen leukergia (ALE) reaction has also been proposed. It is based on the ability of leukocytes of a sensitized organism to agglomerate when exposed to a homologous allergen (dysentery). In view of the lack of evidence of the exact mechanisms of such tests, the insufficient correspondence of their results to the etiology of the disease, these methods, after a short period of their use in the USSR, did not subsequently become widespread.

The detection of Shigella antigens in the body is diagnostically equivalent to the isolation of the pathogen. The main advantages of methods for detecting antigens over bacteriological examination, justifying them clinical application, is the possibility of detecting not only viable microorganisms, but also dead and even destroyed ones, which acquires special meaning when examining patients during or shortly after a course of antibiotic therapy.

One of the best methods for rapid diagnosis of dysentery was the immunofluorescent study of feces (Koons method). The essence of the method lies in the detection of shigella by treating the test material with serum containing specific antibodies labeled with fluorochromes. The combination of labeled antibodies with homologous antigens is accompanied by a specific glow of the complexes detected in a fluorescent microscope. In practice, two main variants of the Koons method are used: direct, in which serum containing labeled antibodies against Shigella antigens is used, and indirect (two-stage) using, at the first stage, serum not labeled with fluorochrome (or globulin fraction of anti-shigella serum). At the second stage, fluorochrome-labeled serum is used against globulins of the anti-shigellosis serum used at the first stage. A comparative study of the diagnostic value of two variants of the immunofluorescent method did not reveal large differences in their specificity and sensitivity. In clinical practice, the use of this method is most effective when examining patients in early dates diseases, as well as in more severe forms of infection. A significant disadvantage of the immunofluorescence method is its lack of specificity. The most important reason insufficient specificity of the immunofluorescence reaction is the antigenic relationship of enterobacteria of different genera. Therefore, this method is considered as indicative in the recognition of shigellosis infection.

Various reactions are used to detect shigella antigens without microscopy. These methods make it possible to detect pathogen antigens in feces in 76.5 - 96.0% of patients with bacteriologically confirmed dysentery, which indicates their rather high sensitivity. It is most advisable to use these methods in the late stages of the disease. The specificity of these diagnostic methods is highly estimated by most authors. However, F.M. Ivanov, who used RSK to detect shigellosis antigens in feces, received positive results when examining healthy people and patients with intestinal infections of other etiologies in 13.6% of cases. According to the author, the use of the method is more appropriate for the detection of specific antigens in the urine, since the frequency of nonspecific positive reactions in the latter case is much lower. The use of various research methods makes it possible to detect Shigella antigens in the urine of the vast majority of patients with bacteriologically confirmed dysentery. The dynamics of excretion of antigens in the urine has some features - the detection of antigenic substances in some cases is possible already from the first days of the disease, but with the greatest frequency and constancy it succeeds on the 10-15th day and even at a later date. According to B.A. Godovanny et al., the proportion of positive urine shigella antigens (RSK) after the 10th day of illness is 77% (the corresponding figure for bacteriological examination of feces is 47%). In connection with this circumstance, the study of urine for the presence of pathogen antigens has the value of a valuable additional method in dysentery, primarily for the purpose of late and retrospective diagnosis.

According to N.M. Nurkina, if the antibody immunoreagent is obtained from polyclonal sera, positive indication results are possible if related antigens are present in the sample. For example, with an erythrocyte diagnosticum from a highly active serum against S.flexneri VI, the S.flexneri I-V antigen is also detected, since the Shigella of both subspecies have a common species antigen. Shigella antigens can be determined during the period of illness both in blood serum and in secretions.

Lee Won Ho et al. it has been shown that the frequency of detection of Shigella antigens and their concentration in blood and urine are higher in the first days of the disease and that the concentration of detected antigens is higher in moderate disease than in mild disease.

CM. Omirbayeva proposed a method for indicating the Shigella antigen, based on the use of formalized erythrocytes as a sorbent for antigens from the studied fecal extract, followed by their agglutination with immune sera. Evaluation of the specificity of this method, in our opinion, needs additional research, since fecal extracts contain significant quantities antigens of other bacteria that are not the causative agent of this intestinal disease.

A number of researchers propose enzyme immunoassay as a method for rapid diagnosis of acute dysentery, which, according to many authors, is considered highly sensitive and highly specific. At the same time, the most high level antigen is found in 1-4 days of illness. Despite the obvious advantages of ELISA, which include high sensitivity, the possibility of strict instrumental quantitative accounting, and the simplicity of setting up the reaction, the widespread use of this method is limited due to the need for special equipment.

Monoclonal antibodies, immunoglobulin fragments, synthetic antibodies, LPS silver staining, and other technological advances are recommended to enhance the sensitivity and specificity of various serological antigen detection methods.

It is often not possible to detect the antigen of an infectious agent even when using highly sensitive reactions to detect the AG of the pathogen in the biological substrates of the body, since a significant part of the antigenic substances, apparently, is in the bioassay in the form of immune complexes in the body. When examining patients with bacteriologically confirmed acute dysentery, positive results of determining the antigen by CSC were noted, according to some reports, only in 18% of cases.

T.V. Remneva et al. propose to use ultrasound to disintegrate antibody complexes with pathogen particles, and then determine the pathogen antigen in CSC in the cold. The method was used to diagnose dysentery; urine samples from patients with acute intestinal infections were used as research material.

The use of the precipitation reaction for antigen detection in acute dysentery is not justified due to its low sensitivity and specificity. We believe that the specificity of any method for indicating Shigella antigens can be significantly increased by using monoclonal antibodies to Shigella.

The coagglutination reaction is also one of the methods for the rapid diagnosis of shigellosis, as well as antigens of pathogens of a number of other infections. With shigellosis, the antigens of pathogens can be determined from the first days of the disease throughout the acute period, as well as within 1-2 weeks after the cessation of bacterial excretion. The advantages of the coagglutination reaction are the ease of making diagnosticums, setting up the reaction, economy, speed, sensitivity, and high specificity.

When conducting diagnostics by determining Shigella antigens from the very beginning of the disease, it is most effective, according to many authors, to examine the feces of patients. With the development of the disease, the possibility of detecting Shigella antigens in urine and saliva decreases, although they are found in faeces with almost the same frequency as at the beginning of the disease. It should be borne in mind that in the first 3-4 days of the disease, feces for antigen are somewhat more efficiently examined in RPHA. In the middle of the disease, RPHA and RNAb are equally effective, and starting from the 7th day, RNAb is more effective in the search for the Shigella antigen. These features are due to the gradual destruction of Shigella cells and their antigens in the patient's intestines during the course of the disease. Shigella antigens excreted in urine are relatively smaller than antigens in faeces. Therefore, it is advisable to examine urine in RNAt. In the urine of women, in contrast to the urine of men, due to probable fecal contamination, Shigella antigens are equally often detected using TPHA and RNAb.

Although the antigen is significantly more often (94.5 - 100%) detected in those samples of feces from which it is possible to isolate Shigella than in those samples from which Shigella is not isolated (61.8 - 75.8%), with parallel bacteriological and serological (for antigen) in the study of fecal samples from patients with dysentery in general, shigella was isolated only from 28.2 - 40.0% of samples, and the antigen was found in 65.9 - 91.5% of samples. It is important to emphasize that the species specificity of the detected antigen always corresponds to the specificity of serum antibodies, the titer of which increases to the maximum in dynamics. When focusing on a conditional diagnostic antibody titer, discrepancies in the specificity of such antibodies and the detected antigen can sometimes be observed. This discrepancy is due to insufficient diagnostic reliability of a single determination of the activity of serum antibodies. In this case etiological diagnosis should be set according to the specificity of the detected antigen.

The PCR method for the task of direct detection of signs of the pathogen is close to the methods of indication of antigens. It allows you to determine the DNA of the pathogen and is based on the principle of natural DNA replication, including the unwinding of the DNA double helix, the divergence of DNA strands and the complementary addition of both. DNA replication may not begin at any point, but only in certain starting blocks - short double-stranded sections. The essence of the method lies in the fact that by marking with such blocks a segment of DNA specific only for a given species (but not for other species), it is possible to repeatedly reproduce (amplify) this particular region. Test systems based on the principle of DNA amplification, in most cases, make it possible to detect bacteria and viruses pathogenic for humans, even in cases where they cannot be detected by other methods. The specificity of PCR test systems (with the correct choice of taxon-specific primers, the exclusion of false positive results and the absence of amplification inhibitors in bioassays) in principle allows you to avoid problems associated with cross-reacting antigens, thereby providing very high specificity. The determination can be carried out directly in clinical material containing a live pathogen. But, despite the fact that the sensitivity of PCR can reach a mathematically possible limit (detection of 1 copy of the DNA template), the method is not used in the practice of diagnosing shigellosis due to its relative high cost.

In wide clinical practice, the most widely used among serological methods of research are methods based on determining the level and dynamics of serum antibodies to the alleged causative agent of the disease.

Some authors have determined antibodies to Shigella in coprofiltrates. Coproantibodies appear much earlier than serum antibodies. The activity of antibodies reaches a maximum at 9-12 days, and by 20-25 days they are usually not detected. R. Laplane et al. suggest that this is due to the destruction of antibodies in the intestine under the action of proteolytic enzymes. Coproantibodies cannot be detected in healthy people.

W. Barksdale et al, T.H. Nikolaev et al. report an increase in the efficiency of deciphering the diagnosis and detecting convalescents by simultaneously determining serum and coproantibodies.

The detection of agglutinins in diagnostic titers is possible with bacteriologically confirmed dysentery only in 23.3% of patients. The limited sensitivity of RA is also manifested in insufficiently high titers of agglutinins detected with its help. There is evidence of unequal sensitivity of RA in various etiological forms of shigellosis infection. According to A.A. Klyucharev, antibodies in a titer of 1:200 and above are detected using RA only in 8.3% of patients with Flexner's dysentery and even more rarely with Sonne's dysentery. Positive results of the reaction are not only more often, but also in higher titers are observed with Flexner I-V and Flexner VI dysentery than with Sonne dysentery. Positive results of RA appear from the end of the first week of the disease and are most often recorded in the second or third week. The first 10 days of illness account for 39.6% of all positive reaction results. According to A.F. Podlevsky et al., agglutinins in diagnostic titers are detected in the first week of the disease in 19% of patients, in the second week - in 25% and in the third - in 33% of patients.

The frequency of positive RA results and the height of titers of antibodies detected with its help are directly dependent on the severity of the course of shigellosis infection. According to V.P. Zubareva, the use of antibiotic therapy does not reduce the frequency of positive RA results, however, when antibiotics are prescribed in the first 3 days of the disease, agglutinins are detected in lower titers.

RA has limited specificity. When examining healthy people, positive results of RA were obtained in 12.7% of cases, in 11.3% of cases group reactions were observed. Due to the antigenic relationship of Flexner I-V and Flexner VI bacteria, cross-reactions are especially often observed in the corresponding etiological forms of shigellosis infection.

With the advent of more advanced methods of serodiagnosis of shigellosis infection, RA has gradually lost its significance. The diagnostic value of the agglutination reaction (“Vidal's dysentery reaction”) (RA) in dysentery is estimated by various researchers ambiguously, however, the results of the work of most authors indicate a limited sensitivity and specificity of this method.

Most often, in order to determine antibodies, an indirect (passive) hemagglutination reaction (RPHA) is used. Detailed studies of the diagnostic value of the passive hemagglutination reaction (RPHA) in shigellosis infection were performed by A.V. Lullu, L. M. Schmuter, T. V. Vlohom and a number of other researchers. Their results allow us to conclude that RPHA is one of the most effective methods for the serological diagnosis of dysentery, although it is not without some common drawbacks inherent in the methods of this group.

A comparative study of sensitivity in dysentery RPHA and agglutination reaction shows a great superiority of the first method. According to A. V. Lullu, the average titers of RPHA in this disease exceed the average titers of RA by 15 times (at the height of the disease by 19-21 times), antibodies in high (1:320 - RPHA) are detected when using 4.5 times more often than in the titer (1:160 when setting up the agglutination reaction). With bacteriologically confirmed acute dysentery, a positive reaction of RPHA in diagnostic titers is noted during examination of 53-80% of patients.

Hemagglutinins are detected from the end of the first week of the disease, the detection frequency and antibody titer increase, reaching a maximum by the end of the second and third weeks, after which their titer gradually decreases.

There is a clear dependence of the frequency of positive results of RPHA and hemagglutinin titers on the severity and nature of the course of shigellosis infection. Relevant studies have shown that with erased and subclinical forms of infection, positive results of RPHA were obtained less frequently than with acute clinically pronounced dysentery (52.9 and 65.0%, respectively), while in titers of 1:200 - 1:400, only 4 responded, 2% of sera (with a clinically pronounced form - 31.2%), and with prolonged and chronic forms, positive results of RPHA were noted in 40.8% of patients, including only 2.0% in a titer of 1:200. There are also reports of different sensitivity of RPHA in certain etiological forms of shigellosis infection. According to L.M. Schmuter, the highest hemagglutinin titers are observed in Sonne dysentery and significantly lower titers in Flexner I-V and Flexner VI dysentery. Antibacterial treatment started in the early stages of the disease, due to a decrease in the duration and intensity of antigenic irritation, can cause the appearance of hemagglutinins in the blood serum in lower titers.

Like the agglutination reaction, RPGA does not always make it possible to accurately recognize the etiological form of shigellosis infection, which is associated with the possibility of group reactions. Cross-reactions are observed mainly in Flexner dysentery - between Flexner I-V and Flexner VI dysentery. Humoral immune response in many patients is poorly expressed. The possibility of cross-agglutination due to common antigens is also not excluded. However, the advantages of this method include the simplicity of setting the reaction, the ability to quickly obtain results, and a relatively high diagnostic efficiency. A significant disadvantage this method is that the diagnosis can be established no earlier than the 5th day of the disease, the maximum diagnostic antibody titers can be determined by the 3rd week of the disease, so the method can be classified as "retrospective".

In order to diagnose dysentery, it is also proposed to determine the level of specific circulating immune complexes represented by the S.sonnei O-antigen, connected to a specific antibody, using an indirect “sandwich version” of enzyme immunoassay due to its high sensitivity and specificity. However, the method is recommended to be used only with 5- days of illness.

In patients with dysentery, from the very beginning of the disease, a specific increase in the bacteriofixing activity of the blood due to the antigen-binding activity of erythrocytes is found. In the first 5 days of AII, the determination of the antigen-binding activity of erythrocytes makes it possible to establish the etiology of the disease in 85-90% of cases. The mechanism of this phenomenon is not well understood. It can be assumed that its basis is the binding by erythrocytes due to their C3v receptors (in primates, including humans) or Fcγ receptors (in other mammals) of the antigen-antibody immune complex.

Among the relatively new methods for recording a specific immune response at the cellular level, attention is drawn to the determination of antigen-binding lymphocytes (ASL) that react with a specific, taxonomically significant antigen. Detection of ASL is carried out by various methods - paired agglutination of lymphocytes with an antigen, immunofluorescence, RIA, adsorption of lymphocytes on antigen-containing columns, adhesion of mononuclear cells on glass capillaries, indirect rosette reaction (RNRO). It should be noted that such highly sensitive methods of ASL registration as ELISA and RIA, adsorption of lymphocytes on antigen-containing columns are technically relatively complex and not always available for wide application. The works of a number of authors have shown the high sensitivity and specificity of PHPR for the detection of ASL in various diseases. A number of researchers have revealed a close relationship between the content of ASL in the blood of patients with various pathologies and forms, the severity and period of the disease, its transition to a protracted or chronic form.

Some authors believe that by determining the level of ASL in the dynamics of the disease, one can judge the effectiveness of the therapy. Most authors believe that if it is successful, the number of ASL falls, and if the effectiveness of treatment is insufficient, an increase or stabilization of this indicator is recorded. It has been reported that ASL can be used to quantify sensitization to tissue, bacterial antigens, and antibiotics, which is important. diagnostic value. The ASL method has been used to a limited extent for diagnosing dysentery.

Opportunity early detection ASL, already in the first days after infection, is very important for early diagnosis and timely treatment, which is necessary for the clinician.

Thus, the data presented in the review show that, given the wide spread of dysentery, insufficient sensitivity, and the late appearance of positive results of many diagnostic methods, it is advisable to develop the diagnostic potential for detecting this infection. The data obtained in many infectious diseases on the high efficiency of the ASL method, the early appearance of its positive result, determine the prospect of studying and applying this method in shigellosis.

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A.M.Sadykova

Dysentery laboratory diagnostics

Tү yin: Zhedel іshek infectionalaryn bakylauda, ​​dysentery naқty diagnostics en özu maselesi bolyp tabylady. Bacterial dysenteric dұrys қoyylғan diagnoses nauқaska vaқytynda em zhүrgіzuge zhane epidemica қarsy sharalardy өtkіzu үshіn manyzdy. Obzordagy kөrsetіlgen mәlіmetter, dysentery ken taraluyn negіzdey otyryp, sezіmtaldyғynyn zhetkіlіksіzdіgі zhane kөp degen diagnosticslyқ аdіsterdіn оң nәtizhesіnің kesh anyқtaluyna baylanysty, wasps infektsionny anyktauda diagnostikalyk potentialdy maksatty turde damytu kerek ekenin korsetedі.

Tү hinds fromө zder: diagnostics, dysentery, antigenbaylanystyrushy adis.

A.M.Sadycova

Laboratory diagnostics of dysentery

Summary: Reliable diagnosis of diarhoea is one of the most important issue to control the accute intestinal infection. Exact diagnosis of bacteriosis diarrhoeas have vitae meaning for correct and accurate treatment of a patient and to take necessary antiepidemic measures as well. The members given in the survey, taking into concideration the widespread diarrhoeia, shows the lack of sensibility and late occurrence of positive results of many diagnostic methods. It is essential aimly to develop the diagnostic potential to design the infection.

keywords: diagnostics, dysentery, antigen binding lymphocytes method.