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

[tetano]

BMC Infect Dis


. 2020 Jul 29;20(1):550.
doi: 10.1186/s12879-020-05277-z.
Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus


Fan Yang ¹ , Lihua Xu ² , Fumin Liu ¹ , Hangping Yao ¹ , Nanping Wu ¹ , Haibo Wu ³



Affiliations

• PMID: 32727378
• PMCID: PMC7391517
• DOI: 10.1186/s12879-020-05277-z

Free PMC article

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.

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