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EvaGreen-based Multiplex Real-time PCR Assay for Rapid Differentiation of Wild-Type and Glycoprotein E-Deleted Bovine Herpesvirus-1 Strains

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Title EvaGreen-based Multiplex Real-time PCR Assay for Rapid Differentiation of Wild-Type and Glycoprotein E-Deleted Bovine Herpesvirus-1 Strains
 
Creator Sachinkumar Shivajirao Pawar
 
Subject EvaGreen,Multiplex,Differentiation,Wild-Type,Bovine Herpesvirus-1
 
Description Not Available
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.
 
Date 2021-05-14T14:27:31Z
2021-05-14T14:27:31Z
2017-10-1
 
Type Research Paper
 
Identifier Pawar SS, Meshram CD, Singh NK, Saini M, Mishra BP, Gupta PK. EvaGreen-based Multiplex Real-time PCR Assay for Rapid Differentiation of Wild-Type and Glycoprotein E-Deleted Bovine Herpesvirus-1 Strains. Anim Biotechnol. 2017 Oct 2;28(4):248-252. doi: 10.1080/10495398.2016.1268620. Epub 2017 Jan 6. P
Not Available
1049-5398
http://krishi.icar.gov.in/jspui/handle/123456789/47020
 
Language English
 
Publisher Not Available