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

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 sub...

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Main Authors: Dewi Noor Hidayati, Eko Agus Srihanto, Tri Untari, Michael Haryadi Wibowo, Koichi Akiyama, Widya Asmara
Format: Article
Language:English
Published: Universitas Gadjah Mada, Yogyakarta 2019-06-01
Series:Indonesian Journal of Biotechnology
Subjects:
Online Access:https://jurnal.ugm.ac.id/ijbiotech/article/view/44298
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author Dewi Noor Hidayati
Eko Agus Srihanto
Tri Untari
Michael Haryadi Wibowo
Koichi Akiyama
Widya Asmara
author_facet Dewi Noor Hidayati
Eko Agus Srihanto
Tri Untari
Michael Haryadi Wibowo
Koichi Akiyama
Widya Asmara
author_sort Dewi Noor Hidayati
collection DOAJ
description 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.
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spelling doaj.art-e0a88a78fdc34cff9b3abeea6cfed8372023-02-06T02:59:29ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412019-06-01241344210.22146/ijbiotech.4429824370The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in IndonesiaDewi Noor Hidayati0Eko Agus Srihanto1Tri Untari2Michael Haryadi Wibowo3Koichi Akiyama4Widya Asmara5Department 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 Indonesia, Jalan Frontage Ahmad Yani, Siwalankerto No.68 – 70, Ketintang, Gayungan, Surabaya, Jawa Timur 60231, IndonesiaVeterinary Disease Investigation Lampung (Balai Veteriner Lampung) Jalan Untung Suropati No. 2, Labuhan Ratu, Kedaton, Bandar Lampung 35142, IndonesiaDepartment of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, IndonesiaDepartment of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jalan Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, IndonesiaAdvanced Research Support Centre (ADRES), Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Jl. Fauna No. 2, Karangmalang, Daerah Istimewa Yogyakarta 55281, IndonesiaConsidering 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.https://jurnal.ugm.ac.id/ijbiotech/article/view/44298bovine herpesvirus-1 (bhv-1)glycoprotein dindonesiainfectious bovine rhinotracheitis (ibr)subtypes
spellingShingle Dewi Noor Hidayati
Eko Agus Srihanto
Tri Untari
Michael Haryadi Wibowo
Koichi Akiyama
Widya Asmara
The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
Indonesian Journal of Biotechnology
bovine herpesvirus-1 (bhv-1)
glycoprotein d
indonesia
infectious bovine rhinotracheitis (ibr)
subtypes
title The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
title_full The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
title_fullStr The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
title_full_unstemmed The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
title_short The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia
title_sort establishment of pcr amplification cloning and sequencing of bovine herpesvirus 1 bhv 1 glycoprotein d gene isolated in indonesia
topic bovine herpesvirus-1 (bhv-1)
glycoprotein d
indonesia
infectious bovine rhinotracheitis (ibr)
subtypes
url https://jurnal.ugm.ac.id/ijbiotech/article/view/44298
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