Archives of Virology

, Volume 163, Issue 6, pp 1671–1675 | Cite as

Establishment of a multiplex real-time RT-PCR assay for rapid identification of H6 subtype avian influenza viruses

  • Fan Yang
  • Haibo Wu
  • Fumin Liu
  • Xiangyun Lu
  • Xiuming Peng
  • Nanping Wu
Brief Report


The H6 subtype avian influenza viruses (AIVs) possess the capacity for zoonotic transmission from avian species to humans. Establishment of a specific, rapid and sensitive method to screen H6 AIVs is necessary. Based on the conserved domain of the matrix and H6 AIV hemagglutinin genes, two TaqMan minor-groove-binder probes and multiplex real-time RT-PCR primers were designed in this study. The multiplex real-time RT-PCR assay developed in this study had high specificity and repeatability and a detection limit of 30 copies per reaction. This rapid diagnostic method will be useful for clinical detection and surveillance of H6 AIVs in China.


Avian influenza virus H6 subtype Detection Rapid Sensitivity 



This study was funded by grants from the National Science Foundation of the People’s Republic of China (81502852), Zhejiang Provincial Natural Science Foundation of China (LY15H190003), and the Independent Task of State Key Laboratory for Diagnosis and Treatment of Infectious Diseases (Nos. 2017ZZ10 and 2018ZZ06).

Compliance with ethical standards

Conflict of interest

The authors declare that they had no conflict of interest.

Supplementary material

705_2018_3773_MOESM1_ESM.jpg (224 kb)
Phylogenetic trees of M and HA genes of the avian influenza viruses used in this study. The tree was created by the maximum-likelihood method and bootstrapped with 1,000 replicates using the MEGA software version 6.0. The H6 avian influenza viruses are indicated by triangles. The scale bar represents the distance unit between sequence pairs (JPEG 223 kb)
705_2018_3773_MOESM2_ESM.jpg (86 kb)
Supplementary material 2 (JPEG 86 kb)
705_2018_3773_MOESM3_ESM.docx (21 kb)
Supplementary material 3 (DOCX 21 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, School of MedicineZhejiang UniversityHangzhouChina

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