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The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia Hidayati, Dewi Noor; Srihanto, Eko Agus; Untari, Tri; Wibowo, Michael Haryadi; Akiyama, Koichi; Asmara, Widya
Indonesian Journal of Biotechnology Article in Press
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (14.17 KB) | DOI: 10.22146/ijbiotech.44298

Abstract

Considering the increasing incidence of infectious bovine rhinotracheitis (IBR) in Indonesia, it is necessary to conduct a more in-depth study of bovine herpesvirus-1 (BHV-1) as the causative agent of IBR disease. Previous research reports indicate that one of the BHV-1 subtypes found in Indonesia is the subtype 1.1. Currently, IBR field case detection in Indonesia still uses the serological method (ELISA) that has the potential to give false positive results and cannot explain the virus subtype. Other detection methods such as viral isolation methods take longer time and require adequate resources. This study aimed to determine the BHV-1 subtypes of Indonesian isolates using molecular techniques. The nested PCR using two pairs of primers has succeeded. The nested PCR using two pairs of primers has succeeded to amplify the glycoprotein D (gD) gene. The gD gene fragment was cloned into the pGEM-T plasmid. The analysis of the gD gene sequence was carried out to determine the BHV-1 character of the Indonesian isolates. The results indicated that the isolates were different from the previous isolates, and had similarities (100%) with subtype 1.2 strain SP1777 and SM023.