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Purpose and areas of usage
Innovative development refers to the field of veterinary medicine. It can be used to diagnose leukosis in cattle in blood samples or other biological fluids.
Gist of development
The method occurring during viable diagnosis of leukosis in cattle by a two - stage polymerase chain reaction (PCR) involves: sampling, DNA extraction from samples, carrying out the first stage of PCR, carrying out the second stage of PCR in real - time, visualization and analysis of results.
Main advantages of development
The second stage of the PCR is in real time. This makes it possible to conduct a multiplex analysis (simultaneously, in one test tube, the detection of two targets). For the second stage of PCR, self - developed primers and fluorescence probes are used for two targets: the ENV of the leukosis prognosis gene and the PRP gene, which is specific to cattle (endogenous control). Endogenous control serves to control the quality and quantity of DNA isolated from blood lymphocytes. It is resulting in the prevention of pseudo - negative results. In addition, the use of real - time PCR does not require separation of amplification products in agarose gel (visualization of the results occurs on the computer monitor). It prevents contamination of the working environment by amplification products and makes the method of diagnosis of leukemia of cattle by two - stage PCR method more secure
The results of the development can be introduced into the practice of veterinary diagnostic laboratories.