Registration certificate No. RZN 2016/3905 dated 04.04.2016

Appointment

The set of reagents "Diagnosticum erythrocyte salmonella Vi-antigenic for RPHA" (SED-Vi) is intended for the detection of antibodies to the Vi-antigen of the causative agent of typhoid in human blood serum by passive hemagglutination reaction (RPHA).

Set characteristic

Operating principle

In the presence of antibodies to the causative agent of typhoid fever, hemagglutination of vi-antigen sensitized chicken erythrocytes is observed, which leads to the formation of an "umbrella" of settled erythrocytes at the bottom of the U-shaped wells of the tablet. In the absence of antibodies to the causative agent of typhoid fever, the settled erythrocytes form a "point".

Set composition

Diagnostic characteristics

Diagnosticum should be agglutinated in RPHA with diagnostic salmonella adsorbed serum, Vi receptor, dry (diluted 1:20), to the titer indicated on the serum label. The conditional level of the diagnostic characteristics of the blood serum of healthy people should be considered a serum dilution not higher than 1:20. The analysis time is 3040 minutes. The kit is designed for the study of 42 blood sera in the screening version or 10 blood sera in the titration version.

Precautions

The kit is for in vitro diagnostic use only. The substances included in the kit are inactivated and safe. When working with the kit, SP 1.3.2322-08 and SanPiN 2.1.7.2790-10 should be observed.

Additional equipment and materials

Equipment, materials, solutions:

• 1-channel pipette dispensers with a variable dosing volume of 5 - 40 μl; 40 - 200 μl; 200 - 1000 μl and 1000 - 5000 μl;
• pipette dispensers 8- or 12-channel with a variable dosage volume of 5 - 40 μl and 40 - 200 μl;
• distilled water (GOST 6709-72).

Analyzed samples

The investigated blood serum samples are stored at a temperature from 2 to 8 ° C for no more than 3 days from the moment of blood sampling. It is allowed to store serum in a frozen state at a temperature not higher than minus 18 ° C for no more than 1 year. Before use, the samples are thawed at a temperature of 16 to 25 ° C and mixed by shaking. Re-freezing is not allowed. Samples with bacterial growth and hemolysis should not be used. Before setting up the reaction, the tested sera are heated at 56 ° C for 30 minutes.

Analysis

Preparation of control diagnostic serum (K +)

Prepare a working solution of diagnostic salmonella adsorbed serum, Vi receptor, dry (dilution 1:20) from 0.3 ml (K +). For this, 0.3 ml of phosphate buffered saline (PBS) is added to the contents of the vial with K +. The remaining amount of serum can be aliquoted and stored frozen at a temperature not exceeding minus 18 ° C for no more than 6 months.

Preparation of diagnosticum of erythrocytic salmonella (EDS)

To prepare a working dilution of a suspension of erythrocyte salmonella diagnosticum, 0.6 ml of distilled water is added to the contents of the vial with dry 6% EDMS and left for hydration for 2 hours at a temperature of 16 to 25 ° C. Then 2.4 ml of phosphate buffer solution (PBS) is added to the solution. The working solution is stored at a temperature of 2 to 8 ° C for no more than 1 month. Freezing is not allowed.

RPHA staging for blood serum screening

Blood sera for screening studies are diluted in the plate wells as follows:

• preliminary dilutions of 1:20 are prepared in the first wells of the plate, first adding 190 μl of the RIP solution, then 10 μl of the studied sera. Each serum is added with a separate tip and carefully pipetted (the color of the solution in the wells after adding the sera should change from blue-violet to green);
• screening dilutions of 1:40 are prepared in the second wells by first adding 25 μl of PBS solution and then 25 μl of previously diluted sera and pipetting carefully.

At each setting of RPHA, it is necessary to conduct a control determination of the K + titer. To do this, 50 μl of PBS solution is added to 8 holes of a long row. Then, 50 μl of the K + working solution (1:20) is added to the first well, carefully pipetted and transferred into the next wells, 50 μl each, receiving 2-fold dilutions from 1:40 to 1: 5120. In another 4 wells, 50 μl of PBS solution is added to control SED for the absence of spontaneous hemagglutination.

In all wells of the plate with screening dilutions of the studied sera (except for the first ones containing RIP) and controls add 25 μl of EDM. Stir the SED suspension in a vial or tray before use! The tablet is thoroughly shaken and left at a temperature of 16 to 25 ° C for 30 to 40 minutes until the erythrocytes in the control have completely settled.

Formulation of RPHA during titration of the studied blood sera

Titration of the studied sera and working solution K + is carried out in short rows of the plate. Another short row is used to control the absence of spontaneous hemagglutination of the EDMS.

In the first wells of short rows for titration of the test serum, 180 μl of the RIP solution is added. All other wells add 50 μl of PBS solution.

In the wells with the RIP solution, add 20 μl of the studied sera (get a dilution of 1:10). Each serum is added with its own tip and pipetted carefully (the color of the solution in the wells should change from blue-violet to green). Then, from the first wells, 50 μl are transferred to the next wells of the rows, receiving two-fold dilutions from 1:20 to 1: 1280. At the end of the titration, the solutions in a volume of 50 μl are removed from the last wells.

At each setting of RPHA, it is necessary to conduct a control determination of the K + titer. To do this, 50 μl of PBS solution is added to 8 holes of a long row. Then, 50 μl of the K + working solution (1:20) is added to the first well, carefully pipetted and transferred into the next wells, 50 μl each, receiving 2-fold dilutions from 1:40 to 1: 5120.

To control the diagnosticum for the absence of spontaneous hemagglutination, 50 μl of PBS solution is added to all wells of a short row.

In all wells (except for the first wells of each row for the studied sera containing RIP) add 25 μl of EDM. Stir the SED suspension in a vial or tray before use! The tablet is thoroughly shaken and left at a temperature of 16 to 25 ° C for 30 to 40 minutes until the erythrocytes in the control have completely settled.

Accounting and interpretation of results

Consideration of the results when screening blood serum

The results are recorded according to the conventional scale of four crosses. The titre of serum is considered to be its dilution, which gives hemagglutination by at least 3 (+++) cross.

• ++++ (4+) - agglutinated erythrocytes form an inverted "umbrella" at the bottom of the hole, its edges fall off;
• +++ (3+) - agglutinated erythrocytes form an inverted "umbrella" at the bottom of the hole, its edges are even;
• ++ (2+) - along with agglutinated erythrocytes at the bottom of the well there is a sediment in the form of a small "ring" of non-agglutinated erythrocytes;
• + (1+) - the majority of erythrocytes are not agglutinated and settle in the form of a small "ring";
• (-) - non-agglutinated erythrocytes form a "point" at the bottom of the well.

A positive result is considered to be hemagglutination of erythrocytes loaded with Vi-antigen by at least 3 crosses (+++).

The quality control of the diagnosticum is 4 wells of the control row, into which only the PBS solution and SED were added. There should be no spontaneous hemagglutination in these wells - the reaction is negative (-). Otherwise, the study should be repeated. If hemagglutination appears during repeated setting, then the drug is not used.

Sera with a negative result should be considered free of antibodies to the Vi-antigen at a diagnostic titer of 1:40 and below.

Sera that give a positive result at a dilution of 1:40 should be retested in the variant with serum titration to establish its titer.

Consideration of results when titrating blood serum

The titre of serum is its dilution, which gives hemagglutination not less than 3 (+++) cross.

The quality control of the diagnosticum is the wells of the row for the SED control. There should be no spontaneous hemagglutination in these wells - the reaction is negative (-). Otherwise, the study should be repeated. If hemagglutination appears during repeated setting, then the drug is not used.

DIAGNOSTICUM (Greek, diagnostikos capable of recognizing) - suspensions of neutralized microorganisms used as antigens for serological reactions. The danger of working with living cultures, their ability to variability, and the presence of broad antigenic connections make it advisable to use D. - more standard and homogeneous preparations containing certain antigenic components.

With the help of D. in reactions of agglutination, passive (indirect) hemagglutination (RPHA), etc., specific antibodies are detected in the sera of humans and animals in order to diagnose and study the body's immune state.

D. is especially widely used in laboratory studies for intestinal infections. However, the generality of the antigenic structure of intestinal bacteria determines the presence of cross-reactions and requires differential detection of antibodies. For this purpose, selective suppression of individual antigens is carried out: with the help of phenol or formalin, they suppress O-agglutinability, as suggested by Felix and Olitsky (A. Felix, L. Olitzki, 1928). By influencing with alcohol according to the method of Wien and Sontag or by heating according to Kauffmann, O-diagnosticums with inactivated flagellar antigen are obtained. Even more promising is the use of mono diagnosticums, the principle of creation of which was developed by F.G.Berngoff (1944). These drugs contain only one antigenic component, and they interact only with certain specific antibodies.

The possibility of preserving the properties of bacterial D. by their lyophilization is shown (see).

D., used for serodiagnosis of various infections, are fundamentally similar, however, certain types of these drugs have a certain specificity.

It is generally accepted that RPHA is more sensitive and specific than bacterial agglutination. Formalinized erythrocytes sensitized with antigens obtained by the methods of Boivin and Westphal are used as antigens in RPHA.

It is also possible to use erythrocyte D. to detect antigen in tissues, discharge of patients, extracts of various substances, etc. In these cases, use erythrocytes sensitized with antibodies - "antibody diagnostics".

Viral D. is used mainly in the reaction of complement fixation (RSK), RTGA, and the neutralization reaction. They are prepared from vaccinated culture liquids processed (inactivated) in various ways.

A brief description of the main D. and the scope of their application are presented in the table.

There are also experimental drugs used in scientific work: erythrocyte colidiagnosticums, diphtheria erythrocytic D., hemagglutinating antigens of parotitis viruses, measles hemagglutinating antigen, etc.

Table. Brief description of the main diagnosticums and the purpose of their application

Bibliography: Zuev A. S. Bacterial vi-diagnosticum for the identification of chronic carriers of typhoid bacteria, Zhurn, micr., Epid, and immun., No. 2, p. 51, 1959, bibliogr .; Zuev AS, Novoselova AI and Likina IV Development of a technique for the production of O-diagnosticums and H-monodiagnostics and their use in serodiagnostics of salmonellosis, ibid., No. 3, p. 42, 1956; Immunology and prevention of intestinal infections, ed. S.I.Didenko, p. 180, M., 1962; Karalnik BV Erythrocytic diagnostics, M., 1976; Guidelines for the microbiological diagnosis of infectious diseases, ed. K. I. Matveeva, p. 172, M., 1973; Subbotina Yu. L. and others. Serological diagnostics of salmonellosis and antigenic connections in reactions with erythrocyte and bacterial O-diagnosticums, Zhurn, micr., Epid, and immun., No. 3, p. 19, 1970, bibliogr.

L.B. Epiphany.

Diagnosticum Vi-antigenic erythrocyte salmonella liquid for RPHA. (Research Institute of Epidemiology and Microbiology named after Pasteur)

Diagnosticum V-antigenic erythrocyte salmonella liquid for RPHA

Reagent kit "Diagnosticum erythrocytic salmonella Vi-antigenic for RPHA" (SED-Vi)

Appointment

The set of reagents is intended for the detection of antibodies to the Vi-antigen of the causative agent of typhoid fever in blood serum in the passive hemagglutination reaction (RPHA).

Method principle

In the presence of antibodies to the Vi-antigen of the causative agent of typhoid, hemagglutination of chicken erythrocytes sensitized by the Vi-antigen is observed. This leads to the formation of an "umbrella" of settled erythrocytes at the bottom of the plate wells with a U-shaped bottom. In the absence of antibodies to the Vi-antigen of the causative agent of typhoid fever, the settled erythrocytes form a "point".

Set characteristic

The set is designed for the study of 42 blood sera in the screening version or 10 blood sera in the titration version.

Precautions

The kit is for in vitro diagnostic use only. The components of the kit are safe, however, the blood serum under study, as well as reagents, equipment and instruments in contact with them, can be potentially infectious. When working with them, the following precautions should be taken:

* work in rubber gloves;

* after completion of the work, the test blood serum samples, reagents and instruments in contact with them, should be disinfected with solutions of 6% hydrogen peroxide or 70% ethyl alcohol, or 3% chloramine B in accordance with SP 1.3.2322-08.

When working with the kit, one should observe the "Rules for the device, safety precautions, industrial sanitation, anti-epidemic regime and personal hygiene when working in laboratories (departments, departments) of sanitary and epidemiological institutions of the system of the USSR Ministry of Health" (Moscow, 1981).

Analysis

Preparation of test samples

The investigated blood serum samples are stored at a temperature from 2 to 8 ° C for no more than 3 days from the moment of blood sampling. It is allowed to store serum in a frozen state at a temperature of minus 18 ° C and below no more than 1 year. Before use, samples are thawed at room temperature and mixed by shaking. Re-freezing is not allowed. Specimens with significant bacterial growth should not be used.

Preparation of control serum (K +)

Prepare a working solution of salmonella serum adsorbed Vi (K +) receptor at a dilution of 1:10. For this, the contents of the vial with K + are dissolved by adding 1 ml of PBS solution. Store (in aliquots of 0.2 ml) frozen at a temperature of minus 18 ° C and below for no more than 6 months.

Preparation of erythrocyte diagnosticum (EDS)

0.6 ml of distilled water is added to the contents of the vial with SED and the mixture is left to hydrate for 2 hours at room temperature. Then 2.4 ml of PBS solution is added to the solution. Store at temperatures from 2 to 8 ° C for no more than 6 months. Freezing is not allowed.

RPHA staging during serum screening

Blood serum for screening study is diluted in paired wells of the plate (two wells are used for each serum) as follows:

* preliminary dilutions of 1:20 are prepared in the first wells, first adding 190 μl of RIP solution to them, and then 10 μl of the studied sera, and mix three times by pipetting (the color of the solutions in the wells after adding the sera should change from blue-violet to green );

* Screening dilutions of 1:40 are prepared in the second wells by first adding 25 μl of PBS solution to them, and then 25 μl of previously diluted sera, and mixing by repeated pipetting.

At each setting of RPHA, it is necessary to conduct a control determination of the K + titer. To do this, 50 μl of PBS solution is added to 8 holes of a long row. Then, 50 μl of working solution K + is added to the first well, mixed by pipetting and transferred into the next wells, 50 μl each, receiving 2-fold dilutions from 1:20 to 1: 2560. In another 4 wells, 50 μl of PBS solution is added to control the SED for the absence of spontaneous agglutination.

In the wells with screening dilutions of the studied sera and controls add 25 μl of EDM. Stir the SED suspension in a vial or tray before use! After making the EDMS, the contents of the wells are mixed by tapping on the edge of the plate. The plate is kept at room temperature for 30-40 minutes.

Consideration of the results when screening blood serum

The results are recorded according to the conventional scale of four crosses:

* ++++ (4+) - erythrocytes form an inverted "umbrella" at the bottom of the hole, its edges fall off;

* +++ (3+) - erythrocytes form an inverted "umbrella" at the bottom of the hole, its edges are even;

* ++ (2+) - erythrocytes form a thin ring at the bottom of the hole;

* + (1+) - erythrocytes form a dense ring or disc at the bottom of the hole;

* (-) - erythrocytes form a point at the bottom of the hole.

A positive result is considered to be hemagglutination not less than 3 (+++) cross.

The quality control of the diagnosticum is 4 wells of the control row, into which only the PBS solution and SED were added. There should be no spontaneous agglutination (-) in these wells. In this case, the study should be repeated. If agglutination appears during re-examination, then the drug is not used.

Sera with a negative result should be considered free of antibodies to the Vi-antigen at a diagnostic titer of 1:40 and below.

Sera that give a positive result at a dilution of 1:40 should be re-examined in the variant with serum titration to establish its titer.

Post-injection of RPHA in blood serum titration

The titration of the studied sera and the K + working solution is carried out in short rows of the plate. Another short row is used to control for the absence of spontaneous agglutination of EDMS.

180 μL of RIP solution is added to the first wells of short rows for titration of the studied sera. All other wells add 50 μl of PBS solution.

To control the diagnosticum for the absence of spontaneous agglutination, 50 μl of PBS solution is added to the wells of a short row.

In the wells with the RIP solution add 20 μl of the studied sera. Each serum is added with an individual tip and the solution in the well is mixed by pipetting three times (the color of the solutions in the wells should change from blue-violet to green).

In the first well of a short row for titrating K +, add 100 μl of the working solution K +, in the remaining wells add 50 μl of the PBS solution.

Then, from the first wells of short rows for the studied sera and control sera, transfer 50 μl to the next wells of the rows, receiving two-fold dilutions from 1:20 to 1: 1280. At the end of the titration, the solutions in a volume of 50 μl are removed from the last wells.

In all wells, except for the first wells of each short row, add 25 μl of SED. Stir the SED suspension in a vial or tray before use! After making the EDMS, the contents of the wells are mixed by tapping on the edge of the plate. The plate is kept at room temperature for 30-40 minutes.

Taking into account the results when titrating blood serum

The titre of serum is its dilution, which gives hemagglutination not less than 3 (+++) cross.

The quality control of the diagnosticum is the short-row wells to control the SED. There should be no spontaneous agglutination (-) in these wells. Otherwise, the study should be repeated with a diagnosticum of another series.

Persons who have detected antibodies to the Vi-antigen in a titer of 1:40 and higher are considered suspicious of chronic carrier of the causative agent of typhoid fever. For the final diagnosis, an in-depth bacteriological examination is necessary.

Transportation and storage conditions, shelf life

* Transportation in accordance with SP.3.3.2.1248-03 at temperatures from 2 to 8 ° C. Short-term (up to 10 days) transportation is allowed at temperatures from 10 to 35 ° C.

* Storage in accordance with SP 3.3.2.1248-03 at temperatures from 2 to 8 ° C.

Expiration date - 1 year. After the expiration date, the reagent kit cannot be used.

The set of reagents Diagnosticum Salmonella VI-antigenic is intended for the detection of specific antibodies to the Salmonella typhus Wi-antigen in the human blood serum in the passive hemagglutination reaction (RPHA).

1. SET CHARACTERISTICS

2.1. The principle of the method.

The active principle of the Diagnosticum Salmonella VI-antigenic is Vi-antigen, fixed on the surface of erythrocytes. When interacting with sera containing antibodies to B-antigen, the phenomenon of erythrocyte agglutination is observed.

2.2. COMPOSITION OF THE KIT

1. ANALYTICAL AND DIAGNOSTIC CHARACTERISTICS.

3.1. Diagnosticum should be agglutinated in RPHA by the diagnostic serum Salmonella adsorbed B and B receptor dry in a dilution of at least 1: 160.

The conditional level of the diagnostic characteristics of the blood serum of healthy people should be considered a serum dilution not higher than 1:20.

3.2. The analysis time is 2 hours.

3.3. The set is designed for 8 definitions.

1. PRECAUTIONS

When working with the kit, one should observe the "Rules for the device, safety precautions, industrial sanitation, anti-epidemic regime and personal hygiene when working in laboratories (departments, departments) of sanitary and epidemiological institutions of the system of the USSR Ministry of Health (M., 1981).

The analyzed sera, as well as the reagents in contact with them, should be considered as potentially infectious, capable of maintaining or transmitting HIV, hepatitis virus or any other causative agent of viral infection for a long time - they should be handled with care:

• work with rubber gloves;
• when pipetting, you must use automatic pipettes;
• at the end of the work, the analyzed sera and the reagents in contact with them, the instruments should be treated with a disinfectant solution
• wipe equipment before and after work with 70% ethyl alcohol.

The analyzed sera should be inactivated at 56 ° C for 30 min.

The diagnostic serum Salmonella adsorbed B and B receptor included in the kit is inactivated.

Objective test results are guaranteed under the following conditions:

• store all reagents of the kit at a temperature from 2 to 8 0 С;
• do not use reagents that have expired;
• do not use kit reagents if their packaging is not labeled accordingly;
• for RPHA use reagents included only in this kit.

Vi-Diagnosticum

Diagnosticum V

Vi-Antigenic

The active principle of the diagnosticum is Vi-antigen, fixed on the surface of erythrocytes. When interacting with sera containing antibodies to B-antigen, the phenomenon of erythrocyte agglutination is observed.

Release form

Available in a set of 1 bottle with diagnosticum - 3 ml, diagnostic serum salmonella adsorbed receptor Vi Dry in the form of lyophilisate from 0.1 ml 1 bottle; 0.9% sodium chloride solution - 2 vials of 8 ml; single-use tablet for immunological reactions - 1 pc.

Composition

Reagents Quantity:

Diagnosticum erythrocyte salmonella B-antigen, which is a 0.75% suspension of formalinized and sensitized B-antigen human erythrocytes of O (I) blood group in phosphate buffer solution (pH-7.2 ± 0.2; concentration - 0.06 mol / L). The preservative is formalin. Homogeneous brown suspension without flakes; when settling, 2 layers are formed: a dense brown precipitate of erythrocytes and a transparent yellowish supernatant liquid 1 bottle-3 ml.

Serum diagnostic salmonella adsorbed B receptor dry - homogeneous mass from white with brownish tint to beige. 1 bottle - from 0.1 ml.

Support solution - 0.9% sodium chloride solution - clear, colorless liquid, pH 6.5 to 7.5. 2 bottles - 8 ml each.

Single-use round-bottom plate for immunological reactions - consists of 8 rows, each of which includes 12 wells with a transparent, colorless, round bottom. 1 PC.

Diagnosticum should be agglutinated in RPHA by the diagnostic serum Salmonella adsorbed B and B receptor dry in a dilution of not less than 1/2 of their titer, but not less than 1/160. The diagnosticum should not be agglutinated by the diagnostic salmonella sera adsorbed dry for RA: O receptor 9 - in a dilution of 1:40 and higher, H receptor d - in a dilution of 1:10 and higher.

Indications for use

Designed to detect specific antibodies to the Salmonella typhus Vi-antigen in the human blood serum in the passive hemagglutination reaction (RPHA).

Dosage regimen and method of administration

Human serum samples are used as analyzed samples.
The analyzed sample is stored under conditions preventing bacterial growth at a temperature of 2 to 8 ° C for no more than 72 hours. Freezing is allowed, frozen analyzed samples should be thawed at room temperature before testing.
The analysis of samples with pronounced hemolysis, bacterial growth, as well as those that have been stored for a long time without freezing or re-frozen is not allowed.

ANALYSIS
Preparation of solutions for RPHA.
Dry open vials with diagnostic salmonella serum adsorbed B and receptor and add 1 ml of the attached 0.9% sodium chloride solution, thus obtaining a dilution of 1:10, which is a working dilution.
An opened vial with the diagnostic serum Salmonella adsorbed Bi receptor dry at a dilution of 1:10 in a closed form can be stored at a temperature of 2 to 8 ° C for one month.
Diagnosticum is ready for use. Before opening the vial with diagnosticum, it is necessary to shake gently until a homogeneous suspension is obtained. It is recommended to repeat shaking during work.
The opened bottle with a closed diagnosticum can be stored at a temperature of 2 to 8 ° C for one month.
0.9% sodium chloride solution. Ready to use.

Conducting RPGA.
When monitoring any amount of analyzed sera, it is mandatory to set 1 series of agglutination with the diagnostic serum Salmonella adsorbed B and dry receptor.
For setting RPHA use a plate for immunological reactions of single use. Prepare two-fold serial dilutions of the analyzed sera in 0.05 ml of the attached 0.9% sodium chloride solution starting from 1:10 to 1: 2560 and 1 series of two-fold serial dilutions of diagnostic salmonella serum adsorbed B receptor dry, starting from 1:10 dilution , up to double the titer indicated on the label of the vials of this serum.
Add 0.025 ml of diagnosticum to each well with serum dilutions.

Mandatory controls are:
1. Control of serum diagnostic salmonella adsorbed receptor
We of dry and analyzed serum, which in a dilution of 1:10 in a volume of 0.05 ml, make
in two control wells.
2. Checking for the absence of spontaneous agglutination of the diagnosticum, for which 0.025 ml of the diagnosticum is added to two holes containing 0.05 ml of 0.9% sodium chloride solution.
The tablet is shaken and placed for 1.5-2.0 hours in a thermostat at a temperature of (37 + 1) ° C.

ACCOUNTING OF RESULTS
The reaction is taken into account according to a four-fold system:
4+ - all erythrocytes are agglutinated and evenly cover the bottom of the well;
3+ - almost all erythrocytes are agglutinated. Against their background, there is an inconspicuous ring of settled non-agglutinated erythrocytes;
3+ - along with a uniform agglutinate at the bottom of the well, there is a precipitate of non-agglutinated erythrocytes in the form of a small "ring" or "button";
1+ - the majority of erythrocytes are non-agglutinated and settled in the form of a small “ring” with smooth edges or “buttons” in the center of the bottom of the well.
(-) - there are no signs of agglutination. Erythrocytes settled in the form of a small "ring" with smooth edges or buttons in the center of the well or the bottom of the test tube.
A positive reaction is considered to be at least 3+.
The results obtained in the RPHA can be considered reliable if the serum of the diagnostic salmonella adsorbed Bi receptor dry 1:10 received a positive result in a dilution of not less than 14 titers, and in 2 wells with the analyzed serum and with the diagnostic serum of the diagnostic salmonella adsorbed receptor Bi dry in dilutions of 1:10 there should be no flakes and sediment; in the wells with 0.9% sodium chloride solution and diagnosticum - the reaction is negative.
The antibody titer of the analyzed serum is considered the last dilution of the serum, which still gives positive agglutination of erythrocytes.
Interpretation of results.
Persons who are found to have antibodies to the Vi-antigen in a dilution of 1:40 and higher are considered suspicious of chronic typhoid bacterial carriage. However, due to the fact that the diagnosis cannot be made only on the basis of serological examination, an in-depth bacteriological examination is necessary.

Precautions for use

Included in the kit is a diagnostic serum Salmonella adsorbed B and a dry inactivated receptor.
The analyzed sera should be inactivated at 56 ° C for 30 minutes.
The analyzed sera, as well as the reagents, equipment and instruments in contact with them, can be potentially infectious material and should be handled with care:
- work with rubber gloves;
- when pipetting, it is necessary to use automatic devices;
- upon completion of work, the analyzed sera and reagents in contact with them, instruments should be treated with a disinfectant solution;
- Before and after work, wipe the equipment with 96% ethyl alcohol.

Objective test results are guaranteed under the following conditions:
- store all reagents of the kit at a temperature of 2 to 8 ° С;
- do not use reagents with an expired date;
- do not use the kit reagents if there is no appropriate marking on their packaging;
- to carry out RPHA use reagents included only in this kit.