Tool for the differential diagnosis of brucellosis and method thereof

 

(57) Abstract:

The tool is designed for the differential diagnosis of brucellosis by means of the formulation of the reaction between the antigen-antibody. Tool for the differential diagnosis of brucellosis contains cells of Brucella in physiological solution, phenol, and optionally sulfuric acid. These components the product contains the following ratio (per 100 ml of physiological solution with the content of dead cells 13 to 14 billion/ml): 10% solution of phenol 0.5 to 0.6 ml, 8 to 12% sulfuric acid solution of 0.6 - 0.8 ml To obtain a tool for the differential diagnosis of brucellosis spend the concentration of dead cells to content 13 - 14 billion in 1 ml of physiological solution. Next, 100 ml of the resulting solution make of 0.5 - 0.6 ml of 10% solution of phenol. The mixture was shaken and incubated for 5 to 6 hours Then to the mixture is added dropwise 0.6 to 0.8 ml of 8 - 12%-aqueous solution of sulfuric acid, shaken for 4 to 5 min and incubated for 4 to 5 days at room temperature. The resulting tool allows differential diagnosis of infected animals, can be used for the given reaction in a sample of whole blood. 2 S. p. f-crystals.

Isobutane in medicine and veterinary medicine. For the diagnosis of brucellosis in humans and animals there are a number of antigens. For example, in medicine use of Hudelson, which tends to nonspecific reactions. In animal brucellosis is a single antigen for RA, RSK and GSK (in vitro method) and rose Bengal antigen (drip method). All existing antigens identify vaccinated and spontaneously infected with brucellosis, differentiating them, they are malespecific, slightly sensitive, labor-intensive, and are intended for the production of the reaction only in the serum.

Closest to the claimed method is a diagnostic tool for the detection of brucellosis, containing killed Brucella cells in finalizando physiological solution [1].

Given all the shortcomings of existing diagnostics, as well as lack of funds for differential diagnosis of vaccinated and infected animals, we have developed a tool and a method thereof, which would meet modern requirements practical Virology. We have developed a diagnostic tool can be successfully used as in blood serum and whole blood, and milk.

Osnovivaite. The initial level of microbial bodies contained in the specified antigen,lead up to number 13 - 14 billion/cm3, then make a 10% solution of phenol, shake the mixture and leave for 5-6 hours then the mixture was added dropwise concentrated sulfuric acid, previously diluted with distilled water in the ratio 1 : 10, shaken for 4 to 5 minutes and leave the mixture for 4 to 5 days, after this time the product is ready to use.

The drug, used for diagnostics, contains a final concentration of the following components (per 100 ml antigen):

Phenol 10% - 0,5 - 0,6 ml

Concentrated sulphuric acid is 0.6 - 0.8 ml

This diagnostic tool used for the detection of brucellosis plate (drip) method).

Diagnostics using the funds obtained as follows.

On the glass cause of 0.03 ml of serum and the top of 0.025 ml, mix thoroughly with a glass rod. Then shake the rotational movements for 5 minutes and take into account the result of the reaction. To accelerate the reaction better after mixing, the mixture was heated to 37 - 40oC. In this case, the result is detected for 3 minutes is athelney - the mixture remains homogeneous, including for vaccinated animals.

Ways of placing reactions in the blood and milk is similar to the method of production of serum.

Control is biofabrics positive hyperimmune serum, diluted 1 : 1 with saline. To verify the specificity and sensitivity of the antigen is taken as 0.03 ml hyperimmune serum and 0.05 ml of antigen, after mixing and rotational movement within 3 min takes into account the result of the reaction. When positive responses are large or small agglutinin in the form of flakes.

The proposed antigen by their sensitivity and specificity allows to determine the antibody titer in the serum drip method. For this purpose, the glass is applied 0,03; 0,02; 0,01; 0,005; 0,0025 ml, brucellosis positive serum, and from above they applied 0,025 ml of antigen. After thorough mixing (which start with a small caption) heated over the flame of a spirit lamp until 37-40oC and by rotational movement in a period of 5 min take into account the results of the reaction. Agglutination in a mixture of 0,03 0,025 corresponds to the title 1 : 100, 0,02 - 0,025 - 1 : 200 and so on

The main advantage is the prob is that when using only whey about 30 40% of the antibodies remain in the blood clot, thus do not allow to identify the disease at an early stage.

This method gives the opportunity by revaccinate to get rid of diseases-human and animal brucellosis, which brings moral and material damage to mankind and livestock.

The preparation method of differential antigen simple, does not require additional investments and changes in production technology biofactories. For setting reactions do not require thermostats and refrigerators.

1. Tool for the differential diagnosis of brucellosis by means of the formulation of the reaction between the antigen antibody containing killed cells of Brucella in physiological solution and phenol, wherein the tool further comprises sulfuric acid in the following ratio of components (per 100 ml of physiological solution with the content of dead cells 13-14 billion/m): a 10% solution of phenol 0,5 0,6 ml, 8-12% solution of sulfuric acid of 0.6 to 0.8 ml.

2. The method of obtaining funds for the differential diagnosis of brucellosis, including the addition of phenol in saline containing killed cells of Brucella, characterized in that conduct kontsentrirovaniem 0,5 0,6 ml of 10% aqueous solution of phenol, the mixture was shaken and incubated for 5 to 6 h, and then to the mixture is added dropwise 0.6 to 0.8 ml 8 12%-aqueous solution of sulfuric acid, shaken for 4 to 5 min and incubated for 4 to 5 days at room temperature.

 

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