【摘要】 目的建立H7N9禽流感病毒一步法逆转录重组酶介导的等温扩增的检测方法(RT-RAA)。方法根据H7N9禽流感病毒HA片段和NA片段的保守区序列分别设计引物、探针,建立RT-RAA方法,研究其特异性、敏感性,并与Realtime RT-PCR方法进行比较。结果成功建立了H7N9禽流感病毒RT-RAA检测方法,H7反应体系和N9反应体系的最低检出限分别为10copy/μL和100copy/μL,且与其他呼吸道病原无交叉反应。H7N9RT-RAA体系检测结果与Real-time RT-PCR一致性较高,Kappa检验u系数为0.933。结论建立的RT-RAA检测方法具有快速、特异及灵敏的特点,为H7N9禽流感病毒的快速检测提供了新的分子工具。更多还原

【Abstract】 Objective To establish a one-step reverse transcription recombinase aided amplification(RT-RAA)assay for the detection of avian-origin influenza A(H7N9)virus.Methods The RT-RAA primers and probes were designed based on conservative regions of haemagglutinin(HA)and neuraminidase(NA)genes of H7N9 virus.Sensitivity and specifi city of the assay were evaluated.The performances of the assay were compared with Real-time RT-PCR using suspected clinical specimens.Results RT-RAA was established for H7N9 detection.The limits of detection for H7N9 were 10copy/μL and 100copy/μL of RNA molecules,respectively.No cross reaction with other pathogens were observed.H7 and N9RT-RAA assay showed good correlation with Real-time RT-PCR assay with Kappa Coefficient uof 0.933 for detection H7N9 for clinical samples.Conclusion The developed RT-RAA assay has characteristics of high sensitivity,specificity and rapidity,which made it a new molecular tool for rapid detection of avian-origin influenza A(H7N9)virus.更多还原