THE POLYMERASE CHAIN REACTION IN THE LABORATORY DIAGNOSTICS OF THE FIRST TYPE HERPESVIRUS INFECTION OF THE HORSES

  • N. Radsikhovskii Zhytomyr National Agroecological University
  • V. Behas Zhytomyr National Agroecological University
  • O. Nikitin Zhytomyr National Agroecological University
Keywords: horses, herpesvirus infections, rhinopneumonia, herpesvirus first type, herpesvirus second type, polymerase chain reaction

Abstract

In Ukraine there is a shortage of domestic, industrial made available diagnostic kits for diagnosis of herpesvirus infections of horses. Some interest is also a comparative assessment of available diagnostic tests for herpes infections of horses. The aim of our research was to develop a method of setting polymerase chain reaction in real time to diagnose herpes infections of horses first type and compare the results obtained with polymerase chain reaction results obtained by other diagnostic tests: haemagglutination inhibition test, agar gel immunodiffusion test. Setting response haemagglutination inhibition test and agar gel immunodiffusion test was carried out in accordance with the guidelines "Diagnosis of herpes infection first and second type of horses." The development of polymerase chain reaction in real time spent at the Laboratory of Quality and Safety of Agricultural Products of the National University of Life and Environmental Sciences of Ukraine. The nucleotide sequence of primers and fluorescent probes to identify horses herpesvirus 1 and 2 types chosen using the program Primer Express (Applied Biosystems). Oligonucleotide primers and fluorescent probes were synthesized amiditophosphit method for automatic DNA synthesizer ABI 3400 (Applied Biosystems). An analysis of nucleotide sequences was found that the most conservative gene in horses herpesvirus types 1 and 2 is a gene encoding the glycoprotein H. We picked the optimal parameters of the polymerase chain reaction to determine the first horse herpesvirus type and analyzed 10 samples of washed lymphocytes. Very high values of limit cycle for these tests Ct 35,5–37,5 indicates a low concentration of the virus in lymphocytes or very low concentration of lymphocytes We also conducted comparing the results obtained in the haemagglutination inhibition test, agar gel immunodiffusion test and polymerase chain reaction. The data indicate a match both positive and negative samples together. Therefore, seronegative animals in agar gel immunodiffusion test and haemagglutination inhibition test can defeat the virus, which in lymphocytes detected by polymerase chain reaction, but not yet developed the disease or those animals are tolerant about the virus. Some of the difference between the response haemagglutination inhibition test and agar gel immunodiffusion test, in our opinion, is that the formation of the infected animals haemagglutinating and precipitating antibodies occurs simultaneously.

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Author Biographies

N. Radsikhovskii, Zhytomyr National Agroecological University
к. вет. наук
V. Behas, Zhytomyr National Agroecological University
к. вет. наук
O. Nikitin, Zhytomyr National Agroecological University
к. вет. наук

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How to Cite
Radsikhovskii, N., Behas, V., & Nikitin, O. (1). THE POLYMERASE CHAIN REACTION IN THE LABORATORY DIAGNOSTICS OF THE FIRST TYPE HERPESVIRUS INFECTION OF THE HORSES. Theoretical and Applied Veterinary Medicine, 5(1), 38-42. Retrieved from https://bulletin-biosafety.com/index.php/journal/article/view/76