Method of preparing diagnostic agglutinating serum for pathogenic yersinia strains

FIELD: medicine.

SUBSTANCE: method of preparing diagnostic agglutinating serum for pathogenic Yersinia strains involves hyperimmunisation of rabbits with antigen representing 0.4-0.6% formalin inactivated antigen (pYV+) of Yersinia enterocolitica My 03R strain into auricular cranial vein. Immunisation of rabbits with said antigen is fourfold in dosage as follows: 290-310 million kl/ml, 490-510 million kl/ml, 0.99-1.1 billion kl/ml and 1.9-2.1 billion kl/ml, respectively with dosage interval 6-7 days. Further the producer is examined for immunogenic properties. Serum separated from the sampled blood is preserved.

EFFECT: method ensures preparation of high-quality agglutinating serum used in yersiniosis diagnostics in animals.

3 ex

 

The invention relates to veterinary medicine and can be used to diagnose yersiniosis and detection of the pathogen in foods of animal and vegetable origin.

A method of obtaining diagnostic agglutinins serum to pathogenic strains of Yersinia by hyperimmunization rabbits antigen of bacteria of the species Yersinia enterocolitica in ear cranial vein, the identification of immunogenic properties of a producer, blood sampling from the producer, followed by the separation and preservation of the target product (Patent of Russia №2203090, MKI AC 39/395, publ. 27.04.03, bull. No. 12).

The disadvantages of this method are the low quality whey, and the danger of its manufacturing process for employees biocarbonate, as in the known method uses a live culture of Yersinia.

Objective of the claimed technical solution is to improve its quality through the use of a new strain of Yersinia and schema changes immunization of rabbits.

The problem is solved in a method of obtaining diagnostic agglutinins serum to pathogenic strains of Yersinia by hyperimmunization rabbits antigen of bacteria of the species Yersinia enterocolitica in ear cranial vein, determine the immunogenic properties of a producer, taking blood from a producer, followed by the separation and preservation target is on the product, that as antigens of bacteria of the species Yersinia enterocolitica using an inactivated 0,4-0,6% formalin antigen plateosauridae (pYV+) strain of Yersinia enterocolitica My 03R, and immunization of rabbits with antigen in the cranial ear vein spend four doses: 290-310 million cells/ml, 490-510 million cells/ml, 0,99-1,1 mlcd cells/ml and 1.9-2.1 billion cells/ml with an interval between each injection 6-7 days.

In the patent and scientific literature is not well-known technical solutions containing modes for diagnostic agglutinins serum to pathogenic strains of Yersinia similarly claimed, i.e. the proposal meets the criterion of "novelty".

All modes method is feasible in laboratory and industrial conditions, aimed at solving real technical problem, i.e. the sentence "industrially applicable".

However, existing serum to pathogenic strains of Yersinia obtained in the process of immunization of animals with antigens of the living culture of Yersinia actually produce without regard to the protective characteristics of the components of the cells of pathogens that are functionally related to pathogenicity factors: protective, surface, soluble antigens of Yersinia and form antibodies to them. The first method for obtaining diagnostic agglutinins serum to pathogenic strains of Yersinia when using the new stamp fee is and Yersinia and schema changes immunization of rabbits, that allows you to get obvious positive effect without compromising the quality of the target product to achieve a faster method of its manufacture, i.e. the proposal meets the criteria of "novelty" and "inventive step".

The method is illustrated by the following examples.

Example 1.

Receive diagnostic agglutinating serum to pathogenic strains of Yersinia as follows. The strain of Yersinia enterocolitica My 03R inactivate 0.4% formalin well-known fashion /F. Kaufman. The family of intestinal bacteria. The pens. from English., Ed. Medgiz, 1959, p.70-150/.

Immunization of rabbits hold antigen in the cranial ear vein four doses: 290 million cells/ml, 490 million cells/ml, 0,99 billion cells/ml and 1.9 billion cells/ml with an interval between each injection 6 days.

7 days after the last injection of antigen in rabbits took the test, got the serum and determined its activity (antibody titer) in RA. The average titer of antibodies in the serum to Yersinia enterocolitica in Armenia amounted to 1:800.

When receiving diagnostic agglutinins serum to pathogenic strains of Yersinia according to known methods (Patent of Russia №2203090, MKI AC 39/395, publ. 27.04.03, bull. No. 12), the mean antibody titer in the serum to Yersinia enterocolitica in RA was 1:400.

Example 2.

Receive diagnostic agglutinating serum to a pathogenic strain is m of Yersinia as follows. The strain of Yersinia enterocolitiea My 03R inactivate 0,6% formalin well-known method.

Immunization of rabbits hold antigen in the cranial ear vein four doses: 310 million cells/ml, 510 million cells/ml, 1,1 mlcd cells/ml and 2.1 billion cells/ml with an interval between each injection 7 days.

6 days after the last injection of antigen in rabbits took the test, got the serum and determined its activity (antibody titer) in RA. The average titer of antibodies in the serum to Yersinia enterocolitica in Armenia amounted to 1:1600.

When receiving diagnostic agglutinins serum to pathogenic strains of Yersinia according to known methods (Patent of Russia №2203090, MKI AC 39/395, publ. 27.04.03, bull. No. 12), the mean antibody titer in the serum to Yersinia enterocolitica in Armenia amounted to 1:800.

Example 3.

Receive diagnostic agglutinating serum to pathogenic strains of Yersinia as follows. The strain of Yersinia enterocolitica My 03R inactivate 0.5% formalin well-known method.

Immunization of rabbits hold antigen in the cranial ear vein four doses: 300 million cells/ml, 500 million cells/ml, 1.0 mlcd cells/ml and 2.0 billion cells/ml with an interval between each injection 6 days.

7 days after the last injection of antigen in rabbits took the test, got the serum and determined its activity (antibody titer) in RA. The average titer obtained in which favorote blood to Yersinia enterocolitica in RA was - 1:1600.

When receiving diagnostic agglutinins serum to pathogenic strains of Yersinia according to known methods (Patent of Russia №2203090, MKI AC 39/395, publ. 27.04.03, bull. No. 12), the average antibody titers in the serum to Yersinia enterocolitica in RA was 1:400.

As shown by experimental results, according to the examples 1-3, the proposed method can improve the quality of the target product 2-4 times and make it safe for workers biological industry.

The method of obtaining diagnostic agglutinins serum to pathogenic strains of Yersinia by hyperimmunization rabbits antigen of bacteria of the species Yersinia enterocolitica in ear cranial vein, the identification of immunogenic properties of a producer, taking blood from a producer, followed by the separation and preservation of the target product, characterized in that as antigens of bacteria of the species Yersinia enterocolitica using an inactivated 0,4-0,6% formalin antigen plateosauridae (pYV+) strain Yersima enterocolitica My 03R, and immunization of rabbits with antigen in the cranial ear vein spend four doses: 290-310 million cells/ml, 490-510 million cells/ml, 0,99-1.1 billion CL/ml and 1.9-2.1 billion cells/ml with an interval between each injection 6-7 days.



 

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