Acta Veterinaria et Zootechnica Sinica ›› 2020, Vol. 51 ›› Issue (6): 1429-1437.doi: 10.11843/j.issn.0366-6964.2020.026

• PREVENTIVE VETERINARY MEDICINE • Previous Articles     Next Articles

Establishment of the Quadruple Real-time Fluorescence RT-PCR for Detection of H5, H7 and H9 Subtypes Avian Influenza Virus

WANG Suchun1, ZHONG Huanxiang2, JIANG Nan3, JIANG Lijian3, PAN Zihao2, SUN Fuliang3, LIU Hualei1, HUANG Baoxu1, WANG Kaicheng1*   

  1. 1. China Animal Health and Epidemiology Center, Qingdao 266032, China;
    2. Nanjing Agricultural University, Nanjing 210095, China;
    3. Yanbian University, Yanji 133000, China
  • Received:2019-11-18 Online:2020-06-25 Published:2020-06-23

Abstract: To shorten the detection time, a quadruple fluorescence RT-PCR method was established for detecting avian influenza virus with the related probes and primers designed and synthesized by analyzing the conservative sequences of M gene, HA gene of H5 subtype, H7 subtype and H9 subtype. This method can detect avian influenza virus, as well as H5 subtype, H7 subtype, and H9 subtype simultaneously. The results showed that the method was quick, specific, and the detection limitation was 10-5 ng·μL-1. Sixty avian influenza viruses were detected from 384 chicken swab samples from 13 living birds markets with the method. All the detected avian influenza viruses were H9 subtype. The results were consistent with the method reported in standard, NY/T 772-2013. The κ value for these two methods was 1 (P<0.001). This method can realize the detection of avian influenza virus which has the characteristics of safety, specificity, rapidity, sensitivity, simplicity and high throughput. It will play an important role in the rapid detection of avian influenza virus.

Key words: avian influenza virus, H5 subtype, H7 subtype, H9 subtype, quadruple real-time fluorescence RT-PCR

CLC Number: