Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus

BMC Infectious Diseases

Lihua Xu¹  Nanping Wu²  Haibo Wu²  Fumin Liu²  Fan Yang²  Hangping Yao² 

[1] Animal Husbandry and Veterinary Institute, Zhejiang Academy of Agricultural Science;State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University;

关键词: Avian influenza virus;    H5N8;    Virus detection;    Minor groove binder probes;    Multiplex real-time RT-PCR;   

DOI  :  10.1186/s12879-020-05277-z

来源: DOAJ

【 摘 要 】

Abstract Background Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. Methods In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. Results The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. Conclusions The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.

【 授权许可】

Unknown