A SYBR Green I-based Quantitative RT-PCR Assay for Bovine Ephemeral Fever Virus and Its Utility for Evaluating Viral Kinetics in Cattle

Overview

Journal J Vet Diagn Invest

Specialty Veterinary Medicine

Date 2019 Dec 18

PMID 31845623

Citations 3

Authors

Shandian Gao

Junzheng Du

Zhancheng Tian

Qingli Niu

Dexuan Huang

Jidong Wang

Jianxun Luo

Guangyuan Liu

Hong Yin

Affiliations

Soon will be listed here.

Abstract

We developed a SYBR green I-based reverse-transcription quantitative PCR (RT-qPCR) assay for bovine ephemeral fever virus (BEFV). Analytical sensitivity of the assay was ~ 100 times higher than conventional RT-PCR. The precision of the RT-qPCR established for RNA standards was high, with intra-assay and inter-assay coefficients of variation of 0.23-0.89% and 0.23-1.02%, respectively. The test was highly specific for BEFV strains, with no cross-reactivity with other viruses of veterinary significance. The assay detected BEFV RNA as early as 1 d post-infection (dpi) and up to 7-8 dpi in the blood samples of experimentally infected cattle. The most stable reference gene, peptidylprolyl isomerase A (), was selected for the quantification of BEFV. Viral RNA loads reached peak level at 3-5 dpi and then decreased rapidly through 7-8 dpi. Our assay provides a reliable approach for the detection of BEFV in the early infection stage and for use in the profiling of BEFV kinetics in vivo.

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