Antigens || Central Research Institute

Proteus Antigens

Description:

Proteus OX2, OX19 & OXK antigen suspensions employed in the Weil-Felix test are used for diagnosis of Rickettsial infection. Rickettisal infection is an arthropod associated infection transmitted by ticks, mites, lice, or fleas to humans as accidental host. Antigen suspension of Proteus OX19 reacts strongly with sera of patients with typhus group and spotted fever group. Proteus OX2 reacts strongly with sera of patients with spotted fever, while the Proteus OXK antigen suspension reacts strongly with sera of patients with scrub typhus.

Product:

Antigen	Proteus OX2	Proteus OX19	Proteus OXK
Packing	50 ml	50 ml	50 ml

Reagents:

Proteus antigens are ready to use smooth antigen suspensions of P.mirabilis strain OXK and Proteus vulgaris strain OX2 and OX 19. These reagents are suitable for use in a standard tube agglutination test procedure for the detection of Rickettsial antibodies in the patient's serum.

Each batch of reagents undergoes quality control testing at various stages of manufacture.

Composition:

The killed suspensions of Proteus mirabilis(OXK),Proteus vulgaris (OX2 and OX 19) strains.
Thiomersal 0.01 % as preservative

Intended use:

Weil Felix test

Principle of the test:

Proteus OX2, Proteus OX19, Proteus OXK antigen suspensions are employed for the Weil-Felix test. The Weil-Felix test is based on the principle that some non-motile strains of Proteus share common somatic antigens with certain species of Rickettsia. Sera from patients infected with Rickettsia will therefore produce agglutination with Proteus antigen suspensions. The smooth, killed Proteus antigen suspensions are mixed with the patient's serum. Antibodies produced due to rickettsial infection if present in the patient serum will react with the Proteus antigen suspension to produce an agglutination reaction. No agglutination indicates the absence of Rickettsial antibodies.

Additional material required:

Round bottom test tubes (12mmx 75 mm), test tube rack, appropriate pipettes/ micropipettes, physiological saline, Water bath (52°C).

Procedure :

Tube agglutination test:

For screening test take 18 round bottom tubes ( 6 each for OX2, OX19& OXK). Prepare doubling dilution of serum from 1:20 to 1:320 and one negative control for OX2, OX19 & OXK each.
Label the tubes as 1-5 for serum dilutions of OX2,OX19 &OXK & 6th as negative control and add 0.5 ml of physiological saline to all tubes.
Prepare 1.5 ml of 1:5 serum dilution by adding 1.2 ml of normal saline & 0.3 ml of serum.
Add 0.5 ml of 1:5 serum dilution to 1^st tube of each OX2,OX19 & OXK, that gives serum dilution of 1:10 in 1^st tube of each OX2,OX19 &OXK.
Now transfer 0.5 ml from 1st tube of OX2, OX19 & OXK to 2^nd, 3^rd, 4^th and 5^th tube and discard 0.5 ml from 5th tube each of OX2, OX19 & OXK.
Now tubes have doubling serum dilutions from 1:10 in 1^st tube to 1:320 in 5^th tube. Now add 0.5 ml of OX2, OX19 & OXK antigens to 1-5^th tubes of 1^st, 2^nd & 3^rd row respectively. That gives us final serum dilutions from 1:20 to 1:320 for OX2, OX19 & OXK respectively.
Incubate the tubes at 52⁰C in water bath for 4 hours.
The control tubes must be examined first and observe the reaction for each antigen.
The positive reaction shows agglutination at the bottom with clearing of suspension.
The negative reaction shows no agglutination..
If negative control shows agglutination reaction then report the test as invalid.
If negative control shows no agglutination then report the test as valid and note the titer of all the tubes.

Warning and Precautions:

These reagents are provided for in vitro diagnostic use only.
Do not use beyond the expiry date.
Wear appropriate protective clothing while handling infectious organisms.
Follow aseptic techniques while testing.
Thiomersal is toxic if ingested.
Do not freeze the antigens.
Only clean and dry test tubes should be used.
Shake vials before use.

Storage:

Proteus antigen should be stored at 2-8ºC and may be used until the expiry date given on the label if handled aseptically.

Expiry:

Shelf life - 9 months from the date of manufacture.

Supply:

Proteus antigen supplied in vials containing 50 ml volume in liquid form.

Salmonella Antigens

Description:

Enteric fever is caused by infection with S. Typhi, S. Paratyphi A and S. Paratyphi B infect the human body .As the response to this infection immune system produce antibodies against these microorganisms . Enteric fever can be diagnosed by Widal test which employs O and H antigen suspension of these microorganisms to detect antibody titer in patient serum. Antibodies to salmonella organisms may be detected in the patient serum from the second week after onset of infection.

Product:

Antigen	S.typhi TO	S. paratyphi AO	S. paratyphi BO	S.typhi TH	S. paratyphi AH	S. paratyphi BH	Salmonella Vi
Packing	50ml	50ml	50ml	50ml	50ml	50ml	50ml

Reagent:

Salmonella antigens are ready to use smooth O & H antigen suspensions of S. typhi , S. paratyphi A and S. paratyphi B. These reagents are suitable for use in a standard tube agglutination test procedure for the detection of S. typhi and S. paratyphi antibodies in the patient's serum.

Each batch of reagents undergoes quality control testing at various stages of manufacture.

Composition:

The killed Salmonella antigen suspensions of S.typhi and S.paratyphi strains.
Thiomersal 0.01 % as preservative

Intended use:

For Widal test

Principle of the test:

When the smooth suspension of Salmonella antigen suspensions are incubated with the patient serum, anti-salmonella antibodies present in the patient's serum react with the antigen suspensions to produce agglutination.

Agglutination is a positive test result, indicating presence of salmonella antibodies in the patient's serum. Absence of agglutination is a negative test result indicating absence of salmonella antibodies in the patient's serum.

Additional material required:

Round bottom felix tubes, Conical bottomed Dreyer’s tubes, patient serum, test tube rack, appropriate pipettes/ micropipettes, physiological saline, Water bath (52°C).

Procedure:

Tube agglutination test:

For screening test take 6 round bottom tubes for S.typhi O antigen in first row and 18 conical bottomed tubes ( 6 each for S. typhi H in 2^nd row, S. paratyphi AH in 3^rd row and S. paratyphi BH antigen in 4^th row).
Prepare doubling dilution of serum from 1:20 to 1:320 and one negative control for S.typhi O, S. typhi H, S. paratyphi AH and S. paratyphi BH each.
Label the tubes as 1-5 for serum dilutions of TO,TH, AH & BH & 6^th as negative control and add 0.5 ml of physiological saline to all tubes.
Prepare 2.0 ml of 1:5 serum dilution by adding 1.6 ml of normal saline & 0.4 ml of serum.
Add 0.5 ml of 1:5 serum dilution to 1st tube of each TO,TH, AH &BH, that gives serum dilution of 1:10 in 1st tube of each TO,TH, AH & BH.
Now transfer 0.5 ml from 1^st tube of TO,TH,AH and BH to 2^nd, 3^rd ,4^th and 5^th tube and discard 0.5 ml from 5^th tube each of TO,TH,AH & BH.
Now tubes have doubling serum dilutions from 1:10 in 1st tube to 1:320 in 5^th tube. Now add 0.5 ml of TO,TH,AH & BH antigens to 1- 5th tubes of 1^st, 2^nd,3^rd and 4^th row respectively. That gives us final serum dilutions from 1:20 to 1:320 for TO,TH,AH & BH respectively.
Add 0.5 ml of antigens of TO,TH, AH & BH to 6^th tube respectively which will act as negative control.
Incubate the tubes at 52⁰C in water bath for 4 hours.
The control tubes must be examined first and observe the reaction for each antigen.
The positive agglutination of O antigen appears as a “matt” or “carpet” at the bottom.
The positive agglutination of H antigen appears as loose, “Wooly” or “Cottony” at the bottom.
The negative agglutination appears as “button” formation at the bottom.
If negative control shows agglutination reaction then report the test as invalid.
If negative control shows no agglutination or “button” formation then report the test as valid and note the titer of all the tubes.