Strain trypanosome trypanosoma equiperdum, ngo "biocenter" how the culture of parasitic protozoa for manufacturer specific trypanosomes of diagnosticum in rac and rdsk

 

(57) Abstract:

The invention relates to the field of veterinary medicine and biotechnology. Strain trypanosome Trypanosoma equiperdum, the registration name of "NGOs "Biocenter", registration number OHMS 1200 used for the manufacture of specific trypanosomes of diagnosticum in RAC and GSK. The strain of Trypanosoma equiperdum (NGOs "Biocenter") is stable, distinct biological, viral, genetic properties, high activity of the antigen, it is well adapted to domestic and farm animals, can cause high parasitemia in the blood of animals. table 1.

The invention relates to the field of agriculture, veterinary Parasitology and biotechnology and can be used to obtain diagnostic specimens for the diagnosis of Trypanosomosis animals: breeding disease (disease one-hoofed), su-aura (disease of cloven-hoofed animals). The strain is used for obtaining a diagnosticum (antigen for binding assays compliment (RAC) and the reaction of long-linking compliment (RDSC)).

Known strains of Trypanosoma allocated by infection of mice with the blood of sick animals (Timofeev B. A., Zablotskiy. Analysis antieau electrophoresis two HSV (variant surface glycoprotein), the clone trypanosome T. Evansi // Chin J. Vet. Sci. 1999, 19 c.3).

The closest in technical essence and the achieved result is a strain of Trypanosoma allocated by infection of mice with the blood of sick animals (Rev. elev. et med vet. Pays trop, 1997, V 50, 1, 29-36).

A common shortcoming of all similar decisions, including the prototype, is the low parasitemia, minor virulent properties, weakly expressed antigenic properties.

The technical result of the present invention is declared strain trypanosome (Trypanosoma equiperdum), with distinct antigenic properties, high parasitemia in animals, and can be used for the preparation of a diagnostic antigen, as well as for diagnostic sera.

The technical result is achieved by the fact that a strain of trypanosome Trypanosoma equiperdum, the registration name of the NGO "Biocenter", registration number MD 12001 as culture parasitic protozoa for manufacturer specific trypanosomes of diagnosticum in RAC and GSK.

The proposed strain of Trypanosoma fusiform shape with a length of 11.9-26,1 mm, a width of 1.8 μm has high activity. Thereto, MOSPRIME the animals I 1 individuals trypanosome. Parasitemia was up to 200 parasites in 1 field of view of the microscope (crushed in a drop of blood taken from infected white mice). Antigen prepared from this strain, reacts with the reference drug (gamma globulin from hyperimmune minimal reverse horses), in breeding 1,50-2 EA (unit activity), and with hyperimmune serum gives 0% hemolysis.

The activity of antigen prepared from this strain is expressed in the marginal and the working title is defined in RAC with hyperimmune and reverses spontaneously patients Trypanosomosis animals, or the reference drug (gamma globulin from hyperimmune serum), respectively, is 1: 200 and 1:50. Strain trypanosome has 8 major polypeptides with m m 35, 44, 47, 50, 54, 71, 86, 92 kDa, of which Supermarine with m m, 44, 54 and 71 kDa.

Strain Trvpanosoma equiperdum (NGOs "Biocenter") has stability, genetic markers, adapted to laboratory animals can cause high parasitemia in the blood. The original strain isolated from a horse, ill breeding disease. After a long long passages on animals (white mice, white rats, Guinea pigs, sheep, dogs) and study its morphological properties, virulence, composition of polipeptides of this strain in RAC with the reference drug, with hyperimmune serum was obtained culture marked NGOs "Biocenter" (, Omsk).

Strain trypanosome Trypanosoma equiperdum NGOs "Biocenter" is characterized by the following properties:

- biological.

Strain well adapted to domestic and farm animals: white mice, white rats, Guinea pigs, dogs, sheep, cattle and horses. Parasitemia was up to 200 parasites in a single field of view of the microscope (crushed in a drop of blood taken from infected white mice). Smear-negative culture, painted with paint from the Institute for Romanovsky, trypanosomes are spindle shaped, with a length of 11.9-26,1 mm, a width of 1.8 μm. The trypanosomes are very active on artificial nutrient media parasites are not cultivated;

- antigenic.

The antigen of this strain of trypanosome has 8 primarnych peptides. Limit and work titer in RAC with hyperimmune serum spontaneously diseased animals, respectively, is 1: 200 and 1: 50 with the reference drug (gamma globulin from hyperimmune serum of horses). Responds in a dilution of 1:50-2 EA (unit activity);

non - virulent.

White mouse killed by injecting from one individual of trypanosome, geneticheskimi characteristics.

Strain trypanosome has 8 major polypeptides with m m 36, 44, 47, 50, 54, 71, 86, 92 kDa, of which supermachine with m m, 44, 54 and 71 kDa;

- range of sensitivity.

Strain Trypanosoma highly sensitive to Nagano, berenil, azedine;

the degree of adaptation and reproduction.

Parasitemia was up to 200 parasites in 1 field of view of the microscope (crushed in a drop of blood from invasioni white mice).

Example.

The study of the biological properties of strains of Trypanosoma (NGOs "Biocenter" , Omsk) compared with strains of France. For these purposes infect white mice parenterally-0.5-0.7 ml blood containing trypanosomes. After 5-7 days bleed and obtained a suspension of blood (0.9% physiological solution of sodium chloride) is administered to dogs (the most convenient model to obtain trypanosomes mass needed in the manufacture of diagnosticum RAC). When reaching the maximum parasites (200 parasites in 1 field of view of the microscope) that is observed at 10-15 day after infection, animals bleed. To the blood to prevent clotting add citric acid sodium or heparin, after which the resulting suspension is centrifuged for 10-15 minutes Ormen elements, and the average white coating on structural elements of trypanosomes.

The top layer is decanted, the layer of trypanosome carefully collected with a spatula. Then they washed initially with the help of 0.9% physiological solution of sodium chloride by double centrifugation for 10 min at 3.5 thousand revs, and then to the precipitate add distilled water for the destruction of Trypanosoma and centrifuged twice for 15 min at 3.5 thousand revs. Subsequently the precipitate washed with a physiological solution of sodium chloride by centrifugation for 20 min at 3.5 thousand revs. The precipitate is destroyed, the trypanosomes. Then these simple is subjected to 3-fold freezing (-15oC) and thawed, and then to add the environment 3 (gelatin, sucrose and distilled water). In the end, this blend is a cytoplasmic antigen used for the diagnosis of Trypanosomosis. The latter using hyperimmune serum or reference serum gamma-globulin from the serum hyperimmune horse) determine the DNC limit and working title activity. As a control using antigen from strain of Trypanosoma received in France. The results are presented Cai get more active antigen for the diagnosis of Trypanosomosis (maximum titer of 1:200, worker 1:50), tested in RAC, compared with strains of France. Thus, the strain GPE "Biocenter" can be used in the production of a specific diagnosis.

Strain trypanosome Trypanosoma equiperdum, the registration name of "NGOs "Biocenter", registration number MD 12001, for the manufacture of specific trypanosomes of diagnosticum in RAC and GSK.

 

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