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One-step triplex reverse-transcription PCR detection of porcine epidemic diarrhea virus, porcine sapelovirus, and porcine sapovirus.

Authors
  • Jiang, Chunyan1, 2
  • He, Haijian1, 2
  • Zhang, Chaoying1, 2
  • Zhang, Xiaoju1, 2
  • Han, Jianfeng1, 2
  • Zhang, Hongbing1, 2
  • Luo, Yu1, 2
  • Wu, Yuan1, 2
  • Wang, Yanli1, 2
  • Ge, Bingqian1, 2
  • Xu, Jia1, 2
  • 1 Department of Animal Production, Faculty of Agriculture and Bioengineering, Jinhua Polytechnic, Jinhua, Zhejiang, China (Jiang, He, C. Zhang, X. Zhang, H. Zhang, Luo, Wu, Wang, Ge, Xu). , (China)
  • 2 Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China (Han). , (China)
Type
Published Article
Journal
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Publication Date
Nov 01, 2019
Volume
31
Issue
6
Pages
909–912
Identifiers
DOI: 10.1177/1040638719883834
PMID: 31650901
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Swine diarrhea can be caused by multiple agents, including porcine epidemic diarrhea virus (PEDV), porcine sapelovirus (PSV), and porcine sapovirus (SaV). We designed a one-step triplex reverse-transcription PCR (RT-PCR) detection method including 3 pairs of primers that focused on the S1 gene of PEDV, a conserved gene of PSV, and the VP1 gene of SaV. The optimal concentrations of upstream and downstream primers in the triplex RT-PCR were 0.24 μM for PEDV, 0.15 μM for PSV, and 0.2 μM for SaV, and the optimal annealing temperature was 55.5°C. Triplex RT-PCR assessment of 402 piglet diarrhea samples was compared with conventional individual RT-PCR. Concordance rates in both tests for individual viruses were 100%, 97.6%, and 94.4% for PEDV, PSV, and SaV, respectively. PEDV, PSV, and SaV were detected in 57.2%, 10.4%, and 9.0% of the samples, respectively. The high sensitivity and specificity of this triplex RT-PCR-based detection method for PEDV, PSV, and SaV could allow rapid detection and analysis of mixed infections by these 3 viruses.

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