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Updated: Jun 20, 2026
Rapid Molecular Detection and Differentiation of Influenza Viruses A and B
Published on: January 30, 2017
[One-step multiplex RT-PCR for rapid screening of type A, B and novel A (H1N1) influenza viruses]
Qiu-hua Mo1, Cui-lan Yang, Ji-can Lin
1Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai 519020, China.
Objective:
To developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.
Methods:
Two pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.
Results:
The RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.
Conclusion:
This multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.
