tetano
Editor, Senior Moderator
J Virus Erad
. 2026 Oct 16;12(4):100638.
doi: 10.1016/j.jve.2026.100638. eCollection 2026 Dec.
Peng Liu 1 2 3 , Xiuwen Zhang 4 5 3 , Yi Wang 2 4 3 , Yan Liang 4 3 , Yangyang Liu 4 3 , Chenyang Wang 1 2 3 , Fei Yu 2 5 3 , Qing'an Han 6 3 , Yuzhong Ma 1
Affiliations Expand
Avian influenza viruses (AIVs) are zoonotic pathogens that exhibit periodic pandemics globally, posing significant public health risks. In this study, we developed a quadruplex RT-qPCR method that targets the M gene for universal AIV detection and the HA gene for simultaneous differentiation of the H5, H7, and H9 subtypes. The assay achieved a detection limit of 1 copy/μL for the H5, H7, and H9 reference materials based on HA gene quantification, equivalent to 5 copies per reaction. No cross-reactivity was observed with a panel of common avian pathogens. The reproducibility evaluation demonstrated both intra-batch and inter-batch coefficients of variation (CVs) to be less than 5%. Furthermore, the assay results were consistent with the expected outcomes for the reference materials tested, and the subtyping results were in agreement with whole-genome sequencing for the environmental samples. These findings indicate that the quadruplex RT-qPCR assay is a reliable tool for broad AIV detection and H5/H7/H9 subtyping, supporting its application in AIV surveillance.
Keywords: Avian influenza virus; Detection; Differentiation; H5/H7/H9 subtypes; Quadruplex RT-qPCR.
. 2026 Oct 16;12(4):100638.
doi: 10.1016/j.jve.2026.100638. eCollection 2026 Dec.
Establishment and performance evaluation of quadruplex RT-qPCR method for universal avian influenza virus detection and H5/H7/H9 subtyping
Peng Liu 1 2 3 , Xiuwen Zhang 4 5 3 , Yi Wang 2 4 3 , Yan Liang 4 3 , Yangyang Liu 4 3 , Chenyang Wang 1 2 3 , Fei Yu 2 5 3 , Qing'an Han 6 3 , Yuzhong Ma 1
Affiliations Expand
- PMID: 42964382
- PMCID: PMC13720467
- DOI: 10.1016/j.jve.2026.100638
Abstract
Avian influenza viruses (AIVs) are zoonotic pathogens that exhibit periodic pandemics globally, posing significant public health risks. In this study, we developed a quadruplex RT-qPCR method that targets the M gene for universal AIV detection and the HA gene for simultaneous differentiation of the H5, H7, and H9 subtypes. The assay achieved a detection limit of 1 copy/μL for the H5, H7, and H9 reference materials based on HA gene quantification, equivalent to 5 copies per reaction. No cross-reactivity was observed with a panel of common avian pathogens. The reproducibility evaluation demonstrated both intra-batch and inter-batch coefficients of variation (CVs) to be less than 5%. Furthermore, the assay results were consistent with the expected outcomes for the reference materials tested, and the subtyping results were in agreement with whole-genome sequencing for the environmental samples. These findings indicate that the quadruplex RT-qPCR assay is a reliable tool for broad AIV detection and H5/H7/H9 subtyping, supporting its application in AIV surveillance.
Keywords: Avian influenza virus; Detection; Differentiation; H5/H7/H9 subtypes; Quadruplex RT-qPCR.