INSTITUTE OF VETERINARY MEDICINE OF THE NAAS OF UKRAINE
发明人:
SYTIUK MYKOLA PETROVYCH,Сытюк Николай Петрович
申请号：
UA201309586
公开号：
UA89920U
申请日:
2013.07.31
申请国别(地区):
UA
年份:
2014
代理人:
摘要：
An immune-peroxidase test for serologic diagnostics of porcine circovirus includes inoculation of the cells of sensitive culture to 96-well plastic microplates with flat bottom dilution of test serums by the method of two-fold dilution with addition of working dilution of circovirus type 2 in the 96-well plastic microplates with round bottom with following contact of those and with bringing to the wells of the microplate with mono-layer of cell culture incubation during seven days under conditions of COincubator removal from the microplate wells of the culture medium, with washing the cell mono-layer introduction of fixing buffer to all the microplate wells step-by-step introduction to all the microplate wells &ndash working dilution of specific blood serum, working dilution of conjugate, solution of substrate termination of reaction through adding stop-solution to each well of the table and registration of reaction under inverted light microscope based on the principle of presence or absence of specific dyeing of the cells &ndash infected cells look like single cells or groups of cells (plaques) with cytoplasm dyed to dark-red color and light nucleus. The test is used for reveal of specific humoral antibodies against the circovirus type 2 in domestic and wild hog through investigation of biological material (serum of blood, milk, colostrums) for cultivation sensitive to circovirus type 2 cell cultures (SK-6, RPK-15, PSGK) are used grown in cultural vessels (glass or plastic pads) cell monolayer is taken by means of solution of trypsin-versen at ratio of 1:9. For cultivation of cell culture, 7-10 % of embryo blood serum of cattle is added to nutrient media the test blood serums before investigation undergo inactivation at temperature during 30 minutes with titration with the method of double dilution with adding working dilution 1000 U of circovirus type 2 (reference - " Stoon 1010" or deposited strains) with exposition during one hour at +. Introduction of content
