Duplex <italic/><i>Taq</i>Man qPCR for Detecting Porcine Epidemic Diarrhea and Transmissible Gastroenteritis Viruses and Epidemic Study in Fujian

CHEN Qiuyong; CHEN Yuanquan; CHEN Rujing; CHE Yongliang; WU Xuemin; LIU Yutao; ZHOU Lunjiang; WANG Longbai

doi:10.19303/j.issn.1008-0384.2022.011.002

Volume 37 Issue 11

Nov. 2022

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Article Navigation > Fujian Journal of Agricultural Sciences > 2022 > 37(11): 1381-1387

CHEN Q Y, CHEN Y Q, CHEN R J, et al. Duplex TaqMan qPCR for Detecting Porcine Epidemic Diarrhea and Transmissible Gastroenteritis Viruses and Epidemic Study in Fujian [J]. Fujian Journal of Agricultural Sciences，2022，37（11）：1381−1387 doi: 10.19303/j.issn.1008-0384.2022.011.002

Citation:

CHEN Q Y, CHEN Y Q, CHEN R J, et al. Duplex TaqMan qPCR for Detecting Porcine Epidemic Diarrhea and Transmissible Gastroenteritis Viruses and Epidemic Study in Fujian [J]. Fujian Journal of Agricultural Sciences，2022，37（11）：1381−1387 doi: 10.19303/j.issn.1008-0384.2022.011.002

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Duplex TaqMan qPCR for Detecting Porcine Epidemic Diarrhea and Transmissible Gastroenteritis Viruses and Epidemic Study in Fujian

doi: 10.19303/j.issn.1008-0384.2022.011.002

1.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences/Fujian Animal Disease Control Technology Development Center, Fuzhou, Fujian　350013, China
2.
Bureau of Animal Husbandry and Veterinary Medicine of Xianyou County, Putian, Fujian　351200, China

Received Date: 2022-08-08
Rev Recd Date: 2022-08-25

Available Online: 2022-12-28

Publish Date: 2022-11-28

Abstract

Abstract

Objective A TaqMan probe-based duplex real-time PCR for rapid detection of porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) was developed. A study was conducted using the methodology to analyze the related 2019–2021 epidemic occurred in Fujian. Method Specific primers and probes labeled with FAM and VIC were designed to amplify the N gene of PEDV and the S gene of TGEV, respectively. A reaction system for the assay was established, optimized, and tested for sensitivity, specificity, and repeatability. The assay was used for the viral detection on 297 suspected clinic specimens collected from 2019 to 2021 for an epidemiology study. Result The newly developed duplex qPCR methodology showed a sensitivity of 10 copies·μL⁻¹ on PEDV and TGEV, which was 100 times higher than that of regular PCR. There were no cross reactions with other common viruses. The inter- and intra-assays had variations on Ct values below 1%. On the 297 specimens, the infection rate of PEDV was 88.89%, that of TGEV 1.01%, and that of both PEDV and TGEV 3.37%. Conclusion The established duplex qPCR had high sensitivity, specificity, repeatability, and reproducibility for detecting PEDV and TGEV. The 2019–2021 epidemic involving the viruses appeared to be mostly PEDV with low incidents of mixed TGEV and PEDV/TGEV infection.
- Porcine epidemic diarrhea virus,
- transmissible gastroenteritis virus,
- duplex real-time PCR,
- epidemiology

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References(21)

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