Acta Veterinaria et Zootechnica Sinica ›› 2023, Vol. 54 ›› Issue (5): 2208-2214.doi: 10.11843/j.issn.0366-6964.2023.05.041

• RESEARCH NOTES • Previous Articles    

Establishment and Preliminary Application of a Real-time RT-RAA for Detection of Transmissible Gastroenteritis Virus

Lü Qiao1,2,3, ZHAO Zhongyi2,3, YIN Dewei2,3, LIU Yumeng2, WANG Wei1, ZHENG Min3, HU Shiyue4, ZHAO Chenchen1, ZHANG Xinyu1, LEI Xiaoxiao1, LU Jingyi1, SUN Wenchao1*, LAN Tian1*   

  1. 1. Wenzhou Key Laboratory for Virology and Immunology, Institute of Virology, Wenzhou University, Wenzhou 325035, China;
    2. College of Animal Science and Technology, Guangxi University, Nanning 530003, China;
    3. Guangxi Center for Animal Disease Prevention and Control, Nanning 530004, China;
    4. Changchun Center for Animal Disease Control and Prevention, Changchun 130061, China
  • Received:2022-06-07 Online:2023-05-23 Published:2023-05-20

Abstract: In order to diagnose transmissible gastroenteritis virus(TGEV)efficiently and easily, specific primers, probes and recombinant standard plasmids TGEV-N were designed. By screening the primers and optimizing the reaction conditions, a real-time reverse-transcription recombinase-aid amplification (RT-RAA) assay was established. The assay can be completed within 20 min at 42℃ and there was no cross-reactivity with porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, porcine deltacoronavirus and porcine enteric alphacoronavirus. Sensitivity test conducted with serial dilutions of TGEV-N ranging from 100-106 copies·μL-1 showed the minimum detection was 6.62×101copies·μL-1. 50 porcine samples were tested by this assay and the positive rate of TGEV was 6%, which was consistent with RT-PCR. The TGEV real-time RT-RAA assay was specific, sensitive and efficient and could be used for clinical screening of TGEV.

Key words: transmissible gastroenteritis virus, real-time RT-RAA, clinical detection

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