Veterynarna biotehnologija – Veterinary biotechnology, 2016, 28, 142-149 [in Ukrainian].
State Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise
VERIFICATION OF ELISA METHOD FOR DIAGNOSIS SPONGIFORM ENCEPHALOPATHY OF CATTLE
Introduction. At the end of the last century, humanity faced the problem of prion infections. Spongiform encephalopathy (SE) in cattle, which in 1985 began to spread massively in the UK, and and later in Europe, causing major economic losses and social disruption.
In 1996, it was found that the BSE pathogen can infect people, causing a new variant Creutzfeldt-Jakob disease. People infected by eating meat products from subclinical sick animals.
Today in the world there is a tendency of reducing the number of cases of SE cow.
For researches on SE of cow in Ukraine it was purchased test kit HerdChek BSE-Scrapie Antigene - for ELISA to detect antigen of bovine spongiform encephalopathy (BSE) - Skrepi (EIA). This test kit was validated in the IDEXX company laboratory.
To establish the possibility of using this test system in a laboratory of the State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise it was necessary to verify the method.
The goal of the work. Verification of ELISA method for the diagnosis of bovine spongiform encephalopathy.
Materials and methods of research. The work was carried out on the basis of research pathomorphological department of the State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise.
For the verification method we used following material:
Interpretation of the results was performed using the «Magellan for F50».
The verification method is performed using a test kit HerdChek BSE-Scrapie Antigen (IDEXX) and equipment for ELISA. The study was conducted according to the guidelines to the test kit.
After the test was carried out calculations of sensitivity, specificity and reproducibility of the method. Determine the coefficient of variation (CV) between holes one plates.
Results of research and discussion. At the first stage of sampling was performed, using disposable syringe 0.30 g were taken (± 0.05 g) of the nervous tissue of the left or right side of the brain stem (obex). Selected tissue was placed in a test tube for crushing and tissue homogenization was performed.
After homogenization of samples brought to the dies, and conducted trials under guidance instructions to the test kit. In the verification method obtained the following results:
In order to monitoring BSE of cattle in Ukraine during 2012, 2013 and 2014 studied 20,061 samples of bovine brain enzyme imunoadsorbtsiyi by using a test set HerdChek BSE-Scrapie Antigen (IDEXX). As a result of investigations the BSE have not found.
Conclusions and prospects for further research:
Keywords: spongiform encephalopathy, cattle, ELISA method, verification.