EvaGreen-based Multiplex Real-time PCR Assay for Rapid Differentiation of Wild-Type and Glycoprotein E-Deleted Bovine Herpesvirus-1 Strains. Issue 4 (2nd October 2017)

• Record Type: Journal Article
• Authors: Pawar, Sachin S.; Meshram, Chetan D.; Singh, Niraj K.; Saini, Mohini; Mishra, B. P.; Gupta, Praveen K.
• Is Part Of: Animal biotechnology. Volume 28:Issue 4(2017)
• Page Start: 248
• Page End: 252
• Publication Date: 2017-10-02
• Subjects: Bovine herpesvirus-1 -- differentiation -- EvaGreen -- wild-type; Animal biotechnology -- Periodicals; Animals, Domestic -- genetics -- Periodicals; Animals, Genetically Modified -- Periodicals; 660.6
• DOI: 10.1080/10495398.2016.1268620
• Languages: English
• ISSNs: 1049-5398

Abstract

ABSTRACT: Bovine herpesvirus-1 (BoHV-1) is an important viral pathogen causing significant economic losses to the cattle industry. Glycoprotein E-deleted marker vaccines form the basis for BoHV-1 control programs widely, wherein detection and differentiation of wild-type and gE-deleted vaccine strains is of crucial importance for proper disease management. In the present study, we report an EvaGreen-based multiplex real-time polymerase chain reaction (EGRT-PCR) assay for rapid differentiation of wild-type and glycoprotein E-deleted strains of BoHV-1. The EGRT-PCR assay could simultaneously detect two viral genes (glycoprotein B and E) and an internal positive control gene (bovine growth hormone- bGH), in a single-tube reaction. The analytical sensitivity of the EGRT-PCR assay was as little as 10 copies of the BoHV-1 DNA per reaction. The modified real-time PCR assay could successfully differentiate wild-type and gE-deleted BoHV-1 strains based on gene specific melting temperatures (Tm ) peaks. Our results have shown that the EGRT-PCR developed in this study might prove to be a promising tool in disease management by enabling rapid differentiation of wild-type and gE-deleted strains of BoHV-1.