【摘要】 [目的 ]　建立应用微流芯片检测甲 1型、甲 3型、乙型流感病毒多重逆转录聚合酶链反应 (mRT -PCR)产物的方法 ,在mRT -PCR扩增核酸的基础上自动化灵敏的定量检测扩增产物 ,快速检测甲亚型、乙型流感病毒 ,帮助临床快速诊断和鉴别诊断其他呼吸道病毒感染 ,以及明确流感病毒在人群中感染、流行情况。　 [方法 ]　采用经MDCK细胞分离培养的甲 1型、甲 3型、乙型流感病毒毒株病毒液 ,使用 3组特异引物经mRT -PCR扩增核酸 ,扩增产物分别采用毛细管电泳技术经Caliper10 0 0微流芯片分析仪自动化检测和经 2 %琼脂糖凝胶电泳法检测。　[结果 ]　设计三组引物对相应甲 1型、甲 3型、乙型流感病毒靶基因的mRT -PCR扩增产物片段分别为 43 0bp、2 10bp、3 91bp ,本实验mRT -PCR扩增产物经 2 %琼脂糖凝胶电泳 ,与Marker比照 ,DNA条带基本在此位点附近 ;产物采用毛细管电泳法经Caliper10 0 0微流芯片分析仪后 ,分别于 413bp、2 0 3bp、3 79bp处出现陡峭的峰 ,如图所示。　[结论 ]　应用微流芯片采用毛细管电泳法可对流感病毒mRT -PCR产物进行定位及相对定量 ,有助于快速诊断流感病毒感染 ,有助于对流感的监测。更多还原

【Abstract】 Objective] To established multiplex reverse transcription-polymerase chain reaction (mRT-PCR) and microchip electrophoresis to detect the influenza A (H1N1 and H3N2) and B viruses in clinical samples and the outbreak and epidemic of influenza in the population. [Methods] Influenza A (H1N1 and H3N2) and B viruses cultured by MDCK shell vial were adopted to detect their DNA zone by agarose gel electrophoresis and microchip electrophoresis after mRT-PCR. [Results] Corresponding obvious DNA zone spectrums at 430bp, 210bp and 391bp respectively in ultraviolet after mRT-PCR and agarose gel electrophoresis; and corresponding DNA peak at 413bp, 203bp and 379bp respectively after mRT-PCR and microchip electrophoresis. [Conclusion] Using three pairs of primers, mRT-PCR and microchip electrophoresis could be used to detect influenza viruses in clinical samples rapidly, which is important for diagnosis and surveillance of influenza.更多还原