Development of a multiplex TaqMan qPCR assay for simultaneous detection and differentiation of four DNA and RNA viruses from clinical samples of sheep and goats.

Authors:
Xingang Xu
Xingang Xu
College of Veterinary Medicine
China
Feng Yang
Feng Yang
West Virginia University
Morgantown | United States
Dr. Qi Zhang, PhD
Dr. Qi Zhang, PhD
Shandong University
Jinan, Shandong | China
Ying Xu
Ying Xu
School of Pharmacy and Pharmaceutical Sciences
San Diego | United States
Jiali Huang
Jiali Huang
National University of Singapore
Singapore
Mingzhe Fu
Mingzhe Fu
College of Animal Science and Technology
Weimin Zhang
Weimin Zhang
School of Life Sciences
China

J Virol Methods 2019 Apr 28;266:58-64. Epub 2019 Jan 28.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China. Electronic address:

Mixed infections with different pathogens are common in sheep and goats under intensive production conditions. Quick and accurate detection and differentiation of different pathogens is necessary for epidemiological surveillance, disease management and import and export controls. Multiplex TaqMan qPCR protocols were developed and subsequently evaluated as effective tools in simultaneously detecting single and mixed infections in sheep and goats. Four pairs of primers and four probes labeled with Rox/BHQ2, Cy5/BHQ2, Hex/BHQ1 and Fam/BHQ1 for peste des petits ruminants virus (PPRV), foot and mouth disease virus (FMDV), goat pox virus (GTPV) and orf virus (ORFV), respectively, were used in the multiplex TaqMan qPCR assay. The assay was shown to be sensitive with detection limits of 9.17 × 10, 1.69 × 10, 9.41 × 10 and 7.46 × 10 copies/μL for PPRV, FMDV, GTPV and ORFV from a mixture of four viruses in a reaction, respectively. The assay was highly specific in its ability to detect one or more viruses in various combinations in the specimens. 38 clinical samples collected from sheep and goats were detected among 43 samples tested by multiplex TaqMan qPCR, showing highly effective identification. Overall, the multiplex TaqMan qPCR panel provides a fast, specific, and sensitive diagnostic tool for the accurate detection of multiple viral pathogens in sheep and goats.

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http://dx.doi.org/10.1016/j.jviromet.2019.01.015DOI Listing
April 2019
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