The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia

Dewi Noor Hidayati (Department of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, Indonesia PUSVETMA (Pusat Veteriner Farma), The Ministry of Agriculture of The Republic of Indones)
Eko Agus Srihanto (Veterinary Disease Investigation Lampung (Balai Veteriner Lampung) Jalan Untung Suropati No. 2, Labuhan Ratu, Kedaton, Bandar Lampung 35142, Indonesia)
Tri Untari (Department of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, Indonesia)
Michael Haryadi Wibowo (Department of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, Indonesia)
Koichi Akiyama (Advanced Research Support Centre (ADRES), Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime 790-8566, Japan)
Widya Asmara (Department of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jl. Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, Indonesia)

Indonesian Journal of Biotechnology
Vol 24, No 1 (2019)

Considering the increasing incidence of infectious bovine rhinotracheitis (IBR) in Indonesia, it was 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 the BHV-1 subtypes found in Indonesia are subtype 1.1. Currently, IBR field case detection in Indonesia still uses the serological method (ELISA), which has the potential to give false positive results and cannot explain the virus subtype. Other detection methods, such as viral isolation, take longer and require adequate resources. This study aimed to determine the BHV-1 subtypes of Indonesian isolates using molecular techniques. Nested PCR using two pairs of primers was successfully used to amplify the glycoprotein D (gD) gene. The gD gene fragment was cloned into the pGEM-T plasmid. Analysis of the gD gene sequence was subsequently 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.