Sinusitis

Diagnosticum vi vi antigenic erythrocyte. Diagnosticum erythrocyte salmonella vi-antigenic

2.diagnosticum

4.from destroyed microbes (isolation of a specific Ag)

Erythrocyte HBs diagnosticum

2.diagnosticum

4. suspension of air-sheep treated with tannin and precipitated a\g HBs

5.to detect specific antibodies in blood serum

Diagnosticum gp120

2.diagnosticum

4.separate a\g ex. HIV

5. for detection of a\t HIV

Tetanus erythrocyte diagnosticum

2.diagnosticum

4. suspension of er.ram, treated with tannin and a\g of tetanus

5. to detect specific antibodies in blood serum (to tetanus

Cardiolipin antigen for microprecipitation

2.diagnosticum

4.extracted lipid fractions from the heart of a healthy bull

5.to detect specific antibodies in blood serum

Ultrasonic treponemal antigen

2.diagnosticum

4. of killed exc. syphilis

5.to detect specific antibodies in blood serum

Corpuscular tularemia diagnosticum

2.diagnosticum

4.from individual particles of the pathogen

5.to detect specific antibodies in blood serum

Dysentery diagnosticum

2.diagnosticum

4. suspension from killed animals

5.to detect specific antibodies in the patient’s blood serum

Erythrocyte diagnosticum from Shigella Sonne

2.diagnosticum

4.suspension of er.ram, treated with tonin

5.to detect specific antibodies in blood serum

Diagnosticum from Salmonella typhimurium

2.diagnosticum

4. of killed exc. salmonella

5.to detect specific antibodies in blood serum

Serums

Anti-gangrenous horse serum 5000 IU

2.serum

4.from the blood serum of hyperimmia horses with an toxoid of excitatory gas gangrene

6. parenterally, after a test with globulin 1:100

Normal human immunoglobulin

2.serum

4.from donor blood serum

5. form-e pass. specific purchased arts imm

6.parenteral

Precipitating anthrax serum

2.serum

4. from the blood serum of hyperimmunized animals a\g exc.sib. ulcers

6.precipitation reaction (RP)

Botulinum serum type – A 400 IU

2.serum

5.to detect a specific antigen in the test material

6.neutralization reaction (RN)

Anti-anthrax globulin

2.serum

4.from the blood serum of hyperimmunized animals and/or exc. anthrax

5. form-e pass. specific purchased arts imm

6.parenteral

Hemolytic serum

2.serum

4.from the serum of hyperimmunized animals with erythrocytes of animals of another type

5.to detect a specific antigen in the test material

Ogawa cholera agglutinating serum

2.serum

4.from blood serum of hyperimmunized animals

5.to detect a specific antigen in the test material

Luminescent tularemia serum

2.serum

4.from the blood serum of hyperimmunized animals a\g exc.tular.

5.to detect a specific antigen in the test material

Immunoglobulin against human tick-borne encephalitis

2.serum

4.from donor blood serum

5. form-e pass. specific purchased arts imm

6.parenteral

ESNO diagnostic serum

2.serum

4.from the blood serum of animals hyperimmunized with ESNO viruses

5.to detect a specific antigen in the test material

Human immunoglobulin against hepatitis B

2.serum

4.from the blood serum of a donor vaccinated against hepatitis B

5. form-e pass. specific purchased arts imm

6.parenteral

Peroxidase-labeled antiglobulin serum

2.serum

4.from blood serum of hyperimmunized animals

5.to detect a specific antigen in the test material (HIV)

Anti-rabies immunoglobulin

2.serum

4.from donor blood serum

5. form-e pass. specific purchased arts imm

The active principle of the diagnosticum is Vi-antigen, fixed on the surface of red blood cells. When interacting with sera containing antibodies to the Vi-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 a lyophilisate of 0.1 ml 1 bottle; 0.9% sodium chloride solution - 2 bottles of 8 ml; single-use tablet for immunological reactions - 1 pc.

Compound

Reagents Quantity:

Diagnosticum erythrocyte Salmonella Vi-antigen, which is a 0.75% suspension of formalinized and sensitized human erythrocytes of blood group O (I) with Vi-antigen in a phosphate buffer solution (pH - 7.2 ± 0.2; concentration - 0.06 mol/l). Preservative - formaldehyde. Homogeneous suspension of brown color without flakes; upon settling, 2 layers are formed: a dense brown sediment of erythrocytes and a transparent yellowish supernatant liquid 1 bottle-3 ml.

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

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

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

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

Indications for use

Designed to detect specific antibodies to Salmonella typhus B antigen in human blood serum in a passive hemagglutination reaction (RPHA).

Dosage regimen and method of administration

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

CONDUCTING THE ANALYSIS
Preparation of solutions for RPGA.
Dry open the vials with diagnostic Salmonella serum adsorbed by the B receptor and add 1 ml of the supplied 0.9% sodium chloride solution, thus obtaining a dilution of 1:10, which is a working dilution.
An opened bottle with Salmonella diagnostic serum adsorbed dry at a dilution of 1:10 can be stored at a temperature of 2 to 8 °C for one month.
Diagnosticum is ready for use. Before opening, the bottle with diagnosticum must be carefully shaken until a homogeneous suspension is obtained. It is recommended to repeat shaking during operation.
An opened bottle with diagnosticum in a closed form 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 number of analyzed sera, it is mandatory to perform 1 series of agglutination with diagnostic Salmonella serum adsorbed by the B receptor dry.
To perform RPHA, a single-use tablet for immunological reactions is used. Prepare two-fold serial dilutions of the analyzed sera in 0.05 ml of the supplied 0.9% sodium chloride solution starting from 1:10 to 1:2560 and 1 row of two-fold serial dilution of diagnostic Salmonella serum adsorbed B receptor dry, starting from a dilution of 1:10 , up to double the titer indicated on the label of the bottles of this serum.
Add 0.025 ml of diagnosticum to each well with serum dilutions.

Mandatory controls are:
1. Control of diagnostic salmonella serum adsorbed by the receptor
Vi dry and analyzed serum, which is added in a dilution of 1:10 in a volume of 0.05 ml
into two control wells.
2. Checking the absence of spontaneous agglutination of the diagnosticum, for which 0.025 ml of diagnosticum is added to two wells 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 using the four-cross system:
4+ - all red blood cells are agglutinated and evenly cover the bottom of the well;
3+ - almost all red blood cells are agglutinated. Against their background there is an inconspicuous ring of settled non-agglutinated erythrocytes;
3+ - along with uniform agglutinate at the bottom of the well there is a sediment of non-agglutinated erythrocytes in the form of a small “ring” or “button”;
1+ - the majority of red blood cells are non-agglutinated and settled in the form of a small “ring” with smooth edges or a “button” in the center of the bottom of the hole.
(-) - there are no signs of agglutination. The red blood cells settled in the form of a small “ring” with smooth edges or a button in the center of the well or the bottom of the test tube.
A reaction of at least 3+ is considered positive.
The results obtained in the RPGA can be considered reliable if the diagnostic Salmonella serum adsorbed B receptor dry 1:10 obtained a positive result in a dilution of not less than 14 of their titers, and in 2 wells with the analyzed serum and with the diagnostic Salmonella serum adsorbed B receptor dry in dilutions of 1:10 there should be no flakes or sediment; in wells with 0.9% sodium chloride solution and diagnosticum - the reaction is negative.
The antibody titer of the analyzed serum is considered to be the last dilution of the serum, which still gives positive agglutination of red blood cells.
Interpretation of results.
Persons who have antibodies to the Vi-antigen at a dilution of 1:40 or higher are considered to be suspected of chronic typhoid bacteria carriage. However, due to the fact that the diagnosis cannot be made only on the basis of a serological study, an in-depth bacteriological examination is necessary.

Precautions for use

Included in the kit are diagnostic serum Salmonella adsorbed B receptor dry inactivated.
Test sera must be inactivated at 56°C for 30 minutes.
Test sera, as well as reagents, equipment and instruments in contact with them, may constitute 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, treat the analyzed sera and reagents and instruments in contact with them with a disinfectant solution;
- wipe equipment with 96% ethyl alcohol before and after use.

Objective analysis results are guaranteed if the following conditions are met:
- all kit reagents should be stored at a temperature of 2 to 8 °C;
- do not use expired reagents;
- do not use the kit reagents if there is no appropriate marking on their packaging;
- to carry out RPGA, use reagents included only in this kit.

The Diagnosticum Salmonella VI-antigen reagent kit is intended for the detection in human blood serum of specific antibodies to the Salmonella typhus VI-antigen in a passive hemagglutination reaction (RPHA).

SET CHARACTERISTICS

2.1. Principle of the method.

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

2.2. SET CONTENTS

Reagents	Quantity
Diagnosticum erythrocyte salmonella Vi-antigenic liquid- is a 1% suspension of formalinized and sensitized Salmonella Typhoid sheep erythrocytes with B-antigen in a phosphate buffer solution (pH 7.2 + 0.2; concentration 0.06 mol/l). Homogeneous suspension of brown color without flakes; upon settling, 2 layers are formed: a dense brown sediment of erythrocytes and a transparent yellowish supernatant liquid	1 bottle - 6 ml
Salmonella diagnostic serum adsorbed B receptor dry - homogeneous mass from white with a brownish tint to beige	1 bottle – 0.1 ml
1% suspension of formalinized, unsensitized sheep erythrocytes— homogeneous suspension of brown color without flakes; upon settling, 2 layers are formed: a dense brown sediment of erythrocytes and a transparent yellowish supernatant liquid	1 bottle –
Solution for diluting serum and staging RPGA - 0.9% sodium chloride solution - transparent colorless liquid, pH from 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 holes with a transparent, colorless, round bottom	1 piece

ANALYTICAL AND DIAGNOSTIC CHARACTERISTICS.

3.1. The diagnosticum must be agglutinated in the RPGA with the diagnostic Salmonella serum adsorbed by the B receptor dry in a dilution of no less than 1:160.

The conditional level of diagnostic characteristics of blood serum from healthy people should be considered a serum dilution of no higher than 1:20.

3.2. Analysis time: 2 hours.

3.3. The set contains 8 definitions.

PRECAUTIONS

When working with the kit, you should follow the “Rules of design, safety precautions, industrial sanitation, anti-epidemic regime and personal hygiene when working in laboratories (departments, departments) of sanitary and epidemiological institutions of the USSR Ministry of Health (M., 1981).

The analyzed sera, as well as reagents in contact with them, should be considered as potentially infected, capable of long-term preservation or transmission of HIV, hepatitis virus or any other causative agent of viral infection - they should be handled with care:

work with rubber gloves;
when pipetting it is necessary to use automatic dispensers;
upon completion of work, treat the analyzed sera and reagents and instruments in contact with them with a disinfectant solution
Wipe equipment with 70% ethyl alcohol before and after use.

The analyzed sera must be inactivated at a temperature of 56 0 C for 30 minutes.

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

Objective analysis results are guaranteed if the following conditions are met:

All kit reagents should be stored at temperatures from 2 to 8 0 C;
do not use expired reagents;
do not use the kit reagents if there is no appropriate marking on their packaging;
To perform RPHA, use reagents included only in this kit.

Vi-Diagnosticum

Diagnosticum Vi

Vi-Antigenic

SET CHARACTERISTICS

2.1. Principle of the method.

2.2. SET CONTENTS

Reagents	Quantity
Diagnosticum erythrocyte salmonella Vi-antigenic liquid- is a 1% suspension of formalinized and sensitized Salmonella Typhoid sheep erythrocytes with B-antigen in a phosphate buffer solution (pH 7.2 + 0.2; concentration 0.06 mol/l). Homogeneous suspension of brown color without flakes; upon settling, 2 layers are formed: a dense brown sediment of erythrocytes and a transparent yellowish supernatant liquid	1 bottle - 3 ml
Salmonella diagnostic serum adsorbed B receptor dry - homogeneous mass from white with a brownish tint to beige	1 bottle – 0.1 ml
1% suspension of formalinized, unsensitized sheep erythrocytes— homogeneous suspension of brown color without flakes; upon settling, 2 layers are formed: a dense brown sediment of erythrocytes and a transparent yellowish supernatant liquid	1 bottle –
Solution for diluting serum and staging RPGA - 0.9% sodium chloride solution - transparent colorless liquid, pH from 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 holes with a transparent, colorless, round bottom	1 piece

ANALYTICAL AND DIAGNOSTIC CHARACTERISTICS.

3.1. The diagnosticum must be agglutinated in the RPGA with the diagnostic Salmonella serum adsorbed by the B receptor dry in a dilution of no less than 1:160.

The conditional level of diagnostic characteristics of blood serum from healthy people should be considered a serum dilution of no higher than 1:20.

3.2. Analysis time: 2 hours.

3.3. The set contains 8 definitions.

PRECAUTIONS

work with rubber gloves;
when pipetting it is necessary to use automatic dispensers;
upon completion of work, treat the analyzed sera and reagents and instruments in contact with them with a disinfectant solution
Wipe equipment with 70% ethyl alcohol before and after use.

The analyzed sera must be inactivated at a temperature of 56 0 C for 30 minutes.

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

Objective analysis results are guaranteed if the following conditions are met:

All kit reagents should be stored at temperatures from 2 to 8 0 C;
do not use expired reagents;
do not use the kit reagents if there is no appropriate marking on their packaging;
To perform RPHA, use reagents included only in this kit.

Registration certificate No. RZN 2016/3905 dated 04/04/2016

Purpose

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

Characteristics of the set

Operating principle

In the presence of antibodies to the causative agent of typhoid fever, hemagglutination of chicken erythrocytes sensitized with the Vi antigen is observed, which leads to the formation of an “umbrella” of settled erythrocytes at the bottom of the U-shaped wells of the plate. In the absence of antibodies to the causative agent of typhoid fever, settled red blood cells form a “dot”.

Set contents

Reagent name	Description	Quantity in set
Diagnosticum erythrocyte salmonella Vi-antigen, dry 6% (EDS)	Formalized chicken erythrocytes sensitized with the S. typhi Vi antigen. Dry hygroscopic mass of brown color. After dissolution, the suspension is red-brown in color.	1 fl., from 0.6 ml
Salmonella diagnostic serum adsorbed, Vi receptor, dry (diluted 1:20, (K+))	Rabbit serum Salmonella adsorbed, Vi receptor, diluted 1:20. Dry hygroscopic porous mass of white color. After dissolution, it is a transparent yellowish or colorless liquid.	1 fl., from 0.3 ml
Test sample diluent (RSD)	Transparent liquid of blue-violet color.	1 fl., 10 ml
Phosphate buffer solution (PBS)	Transparent colorless liquid.	1 fl., 10 ml
Single use polymer tablet for immunological reactions	Single-use polymer tablet for immunological reactions made of transparent, colorless polystyrene.	1 piece

Diagnostic characteristics

The diagnosticum must be agglutinated in the RPHA with the diagnostic Salmonella serum adsorbed, Vi receptor, dry (1:20 dilution), to the titer indicated on the serum label. The conditional level of diagnostic characteristics of blood serum from healthy people should be considered a serum dilution of no higher than 1:20. Analysis time is 3040 minutes. The kit is designed to test 42 blood sera in the screening option or 10 blood sera in the titration option.

Precautions

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

Additional equipment and materials

Equipment, materials, solutions:

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

Analyzed samples

The blood serum samples under study are stored at a temperature of 2 to 8 °C for no more than 3 days from the moment of blood collection. Whey can be stored frozen at a temperature not exceeding minus 18 °C for no more than 1 year. Before use, 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 performing the reaction, the test sera are heated at 56 °C for 30 minutes.

Carrying out analysis

Preparation of control diagnostic serum (K+)

Prepare a working solution of diagnostic Salmonella serum adsorbed, Vi receptor, dry (1:20 dilution) from 0.3 ml (K+). To do this, add 0.3 ml of phosphate buffer solution (PBS) to the contents of the bottle 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 Salmonella erythrocyte diagnosticum (SED)

To prepare a working dilution of the Salmonella erythrocyte diagnosticum suspension, 0.6 ml of distilled water is added to the contents of the bottle with dry 6% SED and left to hydrate 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.

Statement of RPGA during screening of blood serums

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

preliminary dilutions of 1:20 are prepared in the first wells of the plate, first adding 190 μl of RIP solution into them, then 10 μl of the test sera. Each serum is added with a separate tip and carefully pipetted (the color of the solution in the wells after adding the serums 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 into them, and then 25 μl of pre-diluted sera and carefully pipetting.

Each time RPGA is performed, it is necessary to carry out a control determination of the K+ titer. To do this, add 50 μl of PBS solution to 8 wells in a long row. Then add 50 μl of K+ working solution (1:20) to the first well, carefully pipet it and transfer 50 μl to the next wells, obtaining 2-fold dilutions from 1:40 to 1:5120. 50 μl of PBS solution is added to another 4 wells to monitor EDS for the absence of spontaneous hemagglutination.

25 μl of SED is added to all wells of the plate with screening dilutions of the test sera (except for the first ones containing RIP) and controls. Stir the SED suspension in a bottle or bath before use! The tablet is thoroughly shaken and left at a temperature of 16 to 25 ° C for 30 - 40 minutes until the erythrocytes in the control completely settle.

Statement of RPGA during titration of test blood sera

Titration of the test sera and working solution K+ is carried out in short rows of the plate. Another short series is used to monitor the absence of spontaneous hemagglutination of EDS.

180 μl of RIP solution is added to the first wells of short rows for titration of the test serum. Add 50 μl of PBS solution to all other wells.

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

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

25 μl of SED is added to all wells (except for the first wells of each row for test sera containing RIP). Stir the SED suspension in a bottle or bath before use! The tablet is thoroughly shaken and left at a temperature of 16 to 25 ° C for 30 - 40 minutes until the erythrocytes in the control completely settle.

Accounting and interpretation of results

Accounting for results when screening blood serum

The results are taken into account on a conventional scale of four crosses. The serum titer is considered to be its dilution, which gives hemagglutination by at least 3 (+++) crosses.

++++ (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 smooth;
++ (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+) - most red blood cells are not agglutinated and settle in the form of a small “ring”;
(-) - non-agglutinated red blood cells form a “dot” 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 provided by 4 wells of the control row, into which only PBS and SED solution were added. There should be no spontaneous hemagglutination in these wells - the reaction is negative (-). Otherwise, the study should be repeated. If hemagglutination appears upon re-staging, the drug is not used.

Sera with a negative result should be considered not to contain antibodies to the Vi antigen with a diagnostic titer of 1:40 or lower.

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

Accounting for results when titrating blood serum

The serum titer is considered to be its dilution, which gives hemagglutination of at least 3 (+++) crosses.

The quality control of the diagnosticum is provided by the wells of the row for controlling EDS. There should be no spontaneous hemagglutination in these wells - the reaction is negative (-). Otherwise, the study should be repeated. If hemagglutination appears upon re-staging, the drug is not used.