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

• Published in: BMC infectious diseases Vol. 20; no. 1; pp. 1 - 550
• Main Authors: Yang, Fan, Xu, Lihua, Liu, Fumin, Yao, Hangping, Wu, Nanping, Wu, Haibo
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 29.07.2020; BioMed Central; BMC
• Subjects: Analysis; Assaying; Avian flu; Avian influenza; Avian influenza virus; Avian influenza viruses; Deoxyribonucleic acid; DNA; Enzymes; Epidemics; Exo-a-sialidase; Genes; Grooves; H5N8; Hemagglutinins; Infectious diseases; Influenza; Influenza A; Lectins; Minor groove binder probes; Multiplex real-time RT-PCR; Orthomyxoviridae; Plasmids; Polymerase chain reaction; Poultry; Real time; Virus detection; Viruses; Asia; North America; Europe; China; Egypt
• ISSN: 1471-2334; 1471-2334
• DOI: 10.1186/s12879-020-05277-z

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. 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. 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. The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.