Implementation of polymerase chain reaction (PCR) and Real-Time PCR in quick identification of bovine herpesvirus 1

Abstract

Examinations were performed on 65 samples of nasal smeas taken from calves and young cows with clinical symptoms of respiratory infection to determine the presence of the bovine herpes virus 1 using parallel implementation of molecular and standard methods of virological diagnostics. The appearance of a cytopathogenic effect (CPE) was not established in inoculated cell lines 24h, 48h and 72h following inoculation, or after two successive passages of the examined material sample through these cell lines. The application of polymerize chain reaction (PCR) using a primer for glucoprotein B and thymidine - kinasis, established the presence of bovine herpes virus 1 nucleic acid in one sample of a bovine nasal smear, while the presence of this virus was established in three samples in an examination of the nasal smear samples using the Real-Time PCR method.

Ukupno je ispitivano 65 uzoraka nosnih briseva prikupljenih od teladi i junadi sa kliničkim simptomima respiratorne infekcije na prisustvo goveđeg herpesvirusa 1 uporednom primenom molekularnih i standardnih metoda virusološke dijagnostike. Kod inokulisanih ćelijskih linija nije ustanovljena pojava citopatogenog efekta (CPE -) posle 24h, 48h i 72h od inokulacije ni posle dve uzastopne pasaže uzoraka ispitivanog materijala kroz navedene ćelijske linije. Primenom lančane reakcije polimeraze (PCR) uz korišćenje prajmera za glikoprotein B i timidin-kinazu, utvrđeno je prisustvo nukleinske kiseline goveđeg herpesvirusa 1 u jednom uzorku nosnog brisa, dok je ispitivanjem navedenih uzoraka nosnih briseva goveda metodom Real-Time PCR prisustvo pomenutog virusa ustanovljeno kod tri uzorka.