Establishment of a reverse transcription recombinase-aided amplification detection method for porcine group a rotavirus

Yushun Wang; Mincai Nie; Huidan Deng; Siyuan Lai; Yuancheng Zhou; Xiangan Sun; Ling Zhu; Zhiwen Xu

doi:10.3389/fvets.2022.954657

Establishment of a reverse transcription recombinase-aided amplification detection method for porcine group a rotavirus

Front Vet Sci. 2022 Sep 15:9:954657. doi: 10.3389/fvets.2022.954657. eCollection 2022.

Authors

Yushun Wang¹, Mincai Nie¹, Huidan Deng¹, Siyuan Lai¹, Yuancheng Zhou^{2

3}, Xiangan Sun¹, Ling Zhu^{1

4}, Zhiwen Xu^{1

4}

Affiliations

¹ College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
² Animal Breeding and Genetics Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China.
³ Livestock and Poultry Biological Products Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China.
⁴ College of Veterinary Medicine Sichuan Key Laboratory of Animal Epidemic Disease and Human Health, Sichuan Agricultural University, Chengdu, China.

Abstract

Porcine rotavirus type A (PoRVA) is the main cause of dehydration and diarrhea in piglets, which has a great impact on the development of the pig industry worldwide. A rapid, accurate and sensitive detection method is conducive to the monitoring, control, and removal of PoRVA. In this study, a PoRVA real-time fluorescent reverse transcription recombinase-aided amplification (RT-RAA) assay was developed. Based on the PoRVA VP6 gene, specific primers and probes were designed and synthesized. The sensitivity of RT-RAA and TaqMan probe-based RT-qPCR was 7 copies per reaction and 5 copies per reaction, respectively. The sensitivity of the RT-RAA method was close to TaqMan probe-based RT-qPCR. The detection results of RT-RAA and TaqMan probe-based quantitative real-time RT-PCR methods were completely consistent in 241 clinical samples. Therefore, we successfully established a rapid and specific RT-RAA diagnostic method for PoRVA.

Keywords: RT-RAA; RT-qPCR methods; VP6 gene; detection assay; porcine rotavirus.