The strain of bacteria brucella abortus, is used to obtain monospecific serum to identify brucella in r - form

 

(57) Abstract:

The invention is intended for medical immunobiological preparations for the identification of Brucella in the R-form. The strain of bacteria Brucella abortus 3143 is in a stable R-form. It grows well on any nutrient medium used for the cultivation of Brucella. The strain has sufficient immunogenic activity. To obtain serum sufficient single immunization producers (rabbits). The invention will simplify the technology for brucellosis anti-R serum.

The invention relates to medical Microbiology and can be used in obtaining medical immunobiological preparations for the identification of Brucella in the R-form.

As shown by the analysis of known information sources to obtain these drugs are mainly used strains of B. ovis, located in the R-form.

Known reference strain of B. ovis63/290 and strain B/ovic H-61 used as antigens for constructing R-erythrocytic diagnosticums and protivopolozhnyh sera in the R-form (Joint FAO/WHO Expert Committee on Brucellosis. Sixth Report. The World Health Organisation. Technical Report Series, Geneva, 1986. - 133p; Methodical recommendations on preparation and premiu highly hyperimmune protivopolozhnyh sera, Irkutsk, 1988).

The disadvantage of using these strains is the high content of cells in S-shape, which reduces the specificity of the obtained bio-medical drugs to identify R-forms of Brucella.

Known strain of B. ovis 792 (of epidemiology and Microbiology, Moscow) to obtain R-brucellosis diagnostics (RF patent No. 205, 1963, C 12 N1/20, A 61 K 39/10, 10.01.96, Bull. N 1).

The strain of B. ovis 792 isolated from testes of RAM is inputenabled and unagglutinated, infection with this strain of experimental animals causes no formation of antibodies to S-cells of Brucella. The virulence of the strain is low. Antigenic properties are characterized by the formation of specific antibodies after immunization of rabbits.

The disadvantage of using strain is the difficulty of cultivation, due to physiological and biochemical characteristics of the strain. The strain grows in the presence of elevated CO2.

The type strain B. abortus, in particular vaccinal strain B. abortus 19, has a high agglutinogens properties, causes vaccinated animals seropositive reactions that are difficult to distinguish from seropositivity in infected animals.

Known vaccine strain of bacteria B. abort the n against brucellosis. The disadvantage of using strain is the complexity of its selection, which includes five-passage culture through the body of pregnant cows.

The aim of the invention is the finding of a strain which is stable in R-form and with sufficient immunogenic activity when receiving monospecific serum to identify R-forms of Brucella.

This goal is achieved using strain B. abortus 3143 when receiving monospecific serum to identify Brucella in the R-form.

Strain Brucella abortus isolated from an aborted fetus of a cow and is characterized by the following features.

Cultural and morphological characteristics. Cells of B. abortus 3143 represent the cocco-bacteria the size of 0.3-0.6 μm, when the gram stain is negative for painting on Kozlovsky keep the red paint on Romanovsky - pale purple color. Bacteria strains grow well on glucose-glycerol liver broth and agar D, as well as other nutrient media used for cultivation of Brucella and having a neutral or slightly alkaline reaction (pH of 6.8 to 7.2) under aerobic conditions at a temperature of 37oC. Growth in broth with flocculent sludge and t the th form (bipolar), motionless, spores and capsules do not form.

Biochemical characteristics. Culture of bacterial strain B. abortus 3143 is in a stable R-form, while painting on white-Wilson reaction with tripolina positive colonies are red or purple in color, does not produce hydrogen sulfide, grows on environment tinina (2 μg/ml), medium Magenta (1 μg/ml), not agglutinated monospecific sera anti-abortus and anti-melitenzis, agglutinated anti-R serum diagnostic breeding, literoitca phage T6 in RTD-4.

Immunogenic characteristics. The immunization of rabbits causes the formation of specific R-antibodies after a single immunization.

The possibility of practical use of the proposed strain B. abortus 3143 illustrated by examples for diagnostic agglutinins monospecific anti-R serum and serological identification of R-forms of Brucella.

Example 1. A two-day agar culture of the strain B. abortus 3143 plated on the agar plate Albini or meat - peptone-liver agar (MPA) pH of 6.9 to 7.1. After 48 hours incubation at 37oC select colonies and Tsevaot them on sloped agar Albini or MPA in tubes. After 48 hours incubation grown ku is chloride. The obtained suspension was adjusted to a concentration of 20 billion, M. K. ml. of the same solution. Microbial suspension concentration of 20 billion, M. K. injected rabbits weighing 3-3 .5 kg intravenously in a volume of 1.0 ml by the method of Besredka discrete time. After 8-9 days after injection of rabbits totalizer. The resulting serum is used for setting the agglutination reaction.

Example 2. Serum to R-forms of Brucella study strains of Brucella, a typical morphological, cultural and biochemical properties, but in R-form. Agglutination put on the glass. Put a drop agglutinins anti-R serum on glass, and then microbial loop make backass studied the culture of Brucella and suspended smooth stirring. In the formation of agglutinating flakes studied the culture of Brucella defined as the culture of Brucella found in the R-form.

The inventive strain has advantages over the known analogues on the ease of cultivation, high stability of the R-form, sufficient immunogenic, providing the opportunity of single immunization of rabbits to obtain anti-R serum.

The strain of bacteria Brucella abortus 3143 used to obtain monospecific the

 

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