Objective: Developing a rapid method to detect influenza virus N2 subtype by fluorescence real-time quantitative RT-PCR.
Methods: According to conservative sequences of NA gene of influenza virus N2 subtype, a pair primers and Taq-man probe were designed, respectively. Using one step RT-PCR kit to set up real-time RT-PCR system for detection of influenza virus N2 subtype, after the standard quantitative curve of the assay was established using 10-fold serial dilution of TCID50, the sensitivity was determined. The specificity and test for influenza virus illness (ILI) samples were also determined using this RT-PCR system.
Results: The sensitivity of detection influenza virus N2 subtype was 2.56 x 10(-6) and the regression coefficient of the quantitative curve was 0.997. The amplification efficiency and specificity of this assay was 99.9% and 100%, respectively. The real-time RT-PCR for N2 subtype results of ILI samples was in accordance with HAI method completely.
Conclusion: The detection system based on real-time RT-PCR could be utilized to rapidly and sensitively detect influenza virus N2 subtype.