A Taq Man Probe-Based Multiplex Real-Time PCR for Simultaneous Detection of Porcine Epidemic Diarrhea Virus Subtypes G1 and G2, and Porcine Rotavirus Groups A and C

Letian Zhang; Zhiwen Jiang; Zitong Zhou; Jiumeng Sun; Shiyu Yan; Wenting Gao; Yuekun Shao; Yuhe Bai; Yifan Wu; Zefei Yan; Shouzhi Sheng; Alexander Lai; Shuo Su

doi:10.3390/v14081819

A Taq Man Probe-Based Multiplex Real-Time PCR for Simultaneous Detection of Porcine Epidemic Diarrhea Virus Subtypes G1 and G2, and Porcine Rotavirus Groups A and C

Viruses. 2022 Aug 19;14(8):1819. doi: 10.3390/v14081819.

Authors

Letian Zhang¹, Zhiwen Jiang¹, Zitong Zhou¹, Jiumeng Sun¹, Shiyu Yan¹, Wenting Gao¹, Yuekun Shao¹, Yuhe Bai¹, Yifan Wu¹, Zefei Yan¹, Shouzhi Sheng¹, Alexander Lai², Shuo Su¹

Affiliations

¹ Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
² School of Science, Technology, Engineering, and Mathematics, Kentucky State University, Frankfort, KY 40601, USA.

Abstract

Porcine viral diarrhea diseases affect the swine industry, resulting in significant economic losses. Porcine epidemic diarrhea virus (PEDV) genotypes G1 and G2, and groups A and C of the porcine rotavirus, are major etiological agents of severe gastroenteritis and profuse diarrhea, particularly among piglets, with mortality rates of up to 100%. Based on the high prevalence rate and frequent co-infection of PEDV, RVA, and RVC, close monitoring is necessary to avoid greater economic losses. We have developed a multiplex TaqMan probe-based real-time PCR for the rapid simultaneous detection and differentiation of PEDV subtypes G1 and G2, RVA, and RVC. This test is highly sensitive, as the detection limits were 20 and 100 copies/μL for the G1 and G2 subtypes of PEDV, respectively, and 50 copies/μL for RVA and RVC, respectively. Eighty-eight swine clinical samples were used to evaluate this new test. The results were 100% in concordance with the standard methods. Since reassortment between porcine and human rotaviruses has been reported, this multiplex test not only provides a basis for the management of swine diarrheal viruses, but also has the potential to impact public health as well.

Keywords: PEDV G1; PEDV G2; RVA; RVC; detection; real-time PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Coronavirus Infections* / veterinary
Diarrhea / diagnosis
Diarrhea / veterinary
Porcine epidemic diarrhea virus* / genetics
Porcine epidemic diarrhea virus* / isolation & purification
Real-Time Polymerase Chain Reaction / methods
Real-Time Polymerase Chain Reaction / veterinary
Rotavirus* / genetics
Rotavirus* / isolation & purification
Sensitivity and Specificity
Swine
Swine Diseases* / virology

Supplementary concepts

Rotavirus C

Grants and funding

This work was supported by the National Key Research and Development Program of China (grant No. 2021YFD1801101), the Young Top-Notch Talents of the National Ten-Thousand Talent Program, and the Bioinformatics Center (BIC) of Nanjing Agricultural University.