EN - Anglická CZ - Česká SK - Slovenská PL - Polská RO - Rumunská RU - Ruská TR - Turecká UA - Ukrajinská
EN - Anglická
  • CZ - Česká
  • SK - Slovenská
  • PL - Polská
  • RO - Rumunská
  • RU - Ruská
  • TR - Turecká
  • UA - Ukrajinská

Diagnostic kit for farcy using the CBR method

Homepage Products Veterinary products Diagnostic kit for farcy using the CBR method

For serological diagnostics of farcy using complement bond reaction (CBR)

type of preparative: Diagnostics

Can we help you?

Composition:  

The kit contains:

Burkholderia mallei (Pseudomonas mallei) antigen: Transparent, slightly yellowish extract of Burkholderia mallei (Pseudomonas mallei) culture preserved with phenol.

The antigen is calibrated using a standard positive and negative serum, its capacity being adjusted so that in working dilution it provides a visible reaction with positive serum containing a minimum amount of specific antibodies in the dilution of 1:5. In working dilution antigen should not show false positive reactions with negative serum.

Control positive serum: Rabbit serum containing specific complement-fixing antibodies against Burkholderia mallei (Pseudomonas mallei) preserved with phenol.

It is calibrated using a standard antigen its capacity being adjusted so that it exhibits a positive reaction in the dilution 1 : 5 at a minimum.

Haemolytic amboceptor: Rabbit serum containing thermostable antibodies against sheep blood cells preserved with phenol.

Guinea pig complement – guinea pig serum.

Solution for dilution – Solution to re-hydrate the lyophilised antigen and complement to the original volume.

Buffered saline solution according to Mayer-concentrated.

Indication:

For serological diagnostics of farcy using complement bond reaction (CBR).

Performing the test:

Before use antigen and guinea pig complement are diluted to the original volume (1 ml) using a dilution solution that is a part of the kit. For all other dilutions performed during the test Mayer’s concentrated solution is used that is diluted with re-distilled water in the proportion 1 + 4 before use.

Haemolytic amboceptor and complement should be titrated before titration itself.

Titration of haemolytic amboceptor:

Prepare 1:1000 basic dilution and from this an auxiliary dilution series of 1:10,000 or more. Starting with the highest dilution, pipette 0.2 ml of amboceptor of appropriate dilution in test tubes of each of the titration series. Pipette 0.2 ml of 2% suspension of ram blood cells, 0.4 ml of Mayer’s solution, and 0.2 ml of complement diluted 1:20 in all test tubes. At the same time complete the series with three test tubes of common controls: amboceptor, complement, and blood cells. Shake all tubes thoroughly and place immediately in water bath at 37ºC. Read the results after a 30-minute incubation.

Auxiliary dilution of amboceptor                                   

Test tube No.:

1

2

3

4

5

6

7

8

9

10

Final dilution of amboceptor

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

Amboceptor 1:1000 in ml

1

0.5

0.5

0.5

0.5

0.25

0.25

0.25

0.25

0.25

Mayer’s solution ml

-

0.5

1.0

1.5

2.0

1.25

1.5

1.75

2.0

2.25

 

Amboceptor dilution

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

K1

K2

K3

Doses of indiv. dilutions in ml

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

-

-

1 : 100

Mayer’s solution in ml

0.4

0.4

0.4

0.4

0.4

0.4

0.4

0.4

0.4

0.4

0.6

0.6

0.8

Guinea pig complement 1:20

in ml

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

-

0.2

-

2% suspension of sheep blood cells in ml

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

0.2

20-minute incubation in water bath at 37ºC.

K 1 – amboceptor control

K 2 – complement control

K 3 – blood cell control

Evaluation of reaction:

++++                - complete inhibition of haemolysis /+4/

+++, ++, +        - inhibition of haemolysis in gradually decreasing intensity /+3, +2, +1/

-                        - 100% haemolysis – complete lysis of blood cells

Minimum haemolytic dose of amboceptor (MHD) is contained in 0.2 ml of the highest dilution of amboceptor that in the abundance of complement totally dissolves 0.2 ml of the suspension of ram blood cells (the last totally clear test tube). To sensibilise the blood cells, amboceptor is diluted so that it contain 4 MHD in 0.2 ml. The haemolytic system is prepared by mixing 2% suspension of blood cells with the same volume of amboceptor in the working dose of 4 MHD.

Complement titration:

It is diluted immediately before use, using chilled dilution solution, and is stored in the fridge. The basic dilution to proceed from is 1:25. Complement titration is done according to the table.

                                             

Tube No.

1

2

3

4

5

6

7

8

9

10

K1

K2

K3

Complement1 + 24

0.20

0.18

0.16

0.14

0.12

0.10

0.08

0.06

0.04

0.02

0.20

-

-

Mayer’s solution

0.40

0.42

0.44

0.46

0.48

0.50

0.52

0.54

0.56

0.58

0.60

0.60

0.80

Haem. amboceptor 4 MHD

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

-

0.20

-

2% ram blood cells

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

0.20

Incubation in water bath for 30 minutes at 37ºC

% complem.

4%

3.6

3.2

2.8

2.4

2.0

1.6

1.2

0.8

0.4

++++

++++

++++

Results should be read after 30-minute incubation in a water bath at 37ºC. The titre of complement is the smallest amount of complement that, under given conditions, is able to dissolve all blood cells of the haemolytic system. Complement in the dose of 1.5 MHD is used as working dilution (e.g. complement titre is 2.4%, working dilution is 3.6%).

Controls used

K 1 – complement control

K 2 – amboceptor control

K 3 – blood cell control

Examined equine serum should be inactivated before titration in a water bath at 60oC for the period of 20 minutes. Serum of other equidae than horses should be inactivated in a water bath at 63 oC for the period of 35 minutes. Examined serum should be diluted for titration as follows: 1 + 4 (titre 1:5), 1 + 9 (titre 1:10), etc. The same amount of antigen in its working dilution should be added to prepared dilutions of serum followed by the same part of titrated complement (1.5 MHD), and the serum is then incubated in a water bath at 37oC for the period of 30 min. Then a double amount of the haemolytic system is added (the same part of 2% ram blood cells and haemolytic amboceptor - 4 MHD). Incubation is performed in a water bath at 37oC for 30 min. The test for anti-complementarity of the examined serum is performed in the same way. In this series antigen is replaced with Mayer’s dilution solution. A part of titration is represented by control of the guinea pig complement, haemolytic system, and the antigen with control positive serum in its working dilution.

Test serum

Dilution

1:5

1:10

1:20

1:40

-

-

Serum in work. dil.

Dose

0.2

0.2

0.2

0.2

-

-

0.2

-

Antigen

in working dilution

0.2

0.2

0.2

0.2

-

-

0.2

0.2

Complement 1.5 MHD

0.2

0.2

0.2

0.2

0.2

-

0.2

0.2

Incubation at 37ºC for 30 minutes

Haemolytic system

0.4

0.4

0.4

0.4

0.4

0.4

0.4

0.4

Mayer’s solution

 

 

 

 

0.4

0.6

 

0.2

Incubation at 37ºC for 30 minutes

Evaluation of test: 

Evaluation of reaction:

++++                - total inhibition of haemolysis 

+++, ++, +        - inhibition of haemolysis in gradually decreasing intensity

 -                     - complete lysis of blood cells

Sera that in the titre of 1:5 cause inhibition of haemolysis are considered positive.

Sera that in the titre of 1:5 cause a partial inhibition of haemolysis are considered dubious.

Sera that in this titre do not cause any inhibition of haemolysis are considered negative.

In case a dubious result is obtained, the test should be repeated in 2 – 3 weeks.

Notice: Some positive sera may cross-react with Burkholderia pseudomallei (Pseuedomonas pseudomallei).

                                

Storage:

In a dry dark place at the temperature of 2 to 8 °C.
Any packing materials and product residues must be disposed of in accordance with the applicable regulations.

Expirations:

1 year

Notice: Kits expirations is given from complement expiration. It is possible use the others kit components according with their expirations. Guinea pig complement and haemolytic amboceptor are possible to buy separately.

Package:

The kit contains:

Antigen Burkholderia mallei (Pseudomonas mallei)            – 10 ml

Positive control serum                                                          – 1 ml

Haemolytic amboceptor                                                        – 1 ml

Guinea pig complement                                                        – 1 ml

Solution for dilution                                                                – 2 ml

Buffered saline solution according to Mayer –concentrated –100 ml                   


Contact us


antispam
Please fill in the text displayed in the picture

or call


+420 517 318 598