6种核酸提取试剂盒对甲型H1N1流感病毒核酸提取效果的比较

A comparison of six RNA extraction kits for extraction of influenza A/H1N1 virus nucleic acids

目的研究5种国产病毒RNA提取试剂盒对甲型H1N1流感病毒检测的相对效率、耗时和成本,为各级实验室甲型H1N1流感病毒核酸提取检测提供技术参考。方法对滴度为105pfu/ml的甲型H1N1流感病毒参考株悬液作10-1~10-4系列稀释,采用德国Qiagen公司的QIAamp Viral RNAMini Kits和5种国产试剂盒对上述样本进行核酸提取,用复合探针实时荧光定量逆转录聚合酶链反应（RT-PCR）对其提取效率进行平行检测,分析5种国产试剂盒与QIAamp Viral RNA Mini Kits提取效率的差异,同时对试剂盒提取时间及价格进行比较。结果对不同稀释度流感病毒样本的提取检测,各试剂盒存在显著差异,均以QIAamp Viral RNA Mini Kits提取效率最高。综合提取效率、耗时及成本等因素考虑,B试剂盒性价比最高。结论不同核酸提取试剂盒对病毒核酸的提取,可影响RT-PCR的检测效果,各实验室应根据自身条件及试验目的,合理选择性价比较高的试剂盒对临床样本进行核酸提取。

Objective To study the relative efficiency,time-consumption and cost of five domestic viral RNA extraction kits for detection of influenza A/H1N1 viruses,and provide technical reference for relevant laboratories.Methods A suspension containing influenza A/H1N1 viruses 105 pfu/ml was diluted serially from 10-1 to 10-4.RNA extraction kits manufactured by six different companies,including the German Qiagen＇s QIAamp Viral RNA Mini Kits and five types of domestic products were used.Viral RNA extraction was performed based on the instruction of the manufacturers.The RNA preparations were quantified by the complex probe fluorescence real-time PCR.Results The extraction efficiency of six RNA extraction methods had statistically significant difference in the detection of differently diluted flu viruses.Qiagen＇s QIAamp Viral RNA Mini kits gave the highest efficiency.Considering extraction efficiency,time-consumption and cost,Kit B showed the highest cost-effective ratio.Conclusion The results showed that the viral RNA extraction kits could influence obviously the efficiency of real-time PCR assay for influenza A/H1N1 viruses.So the choice of kits for viral RNA extraction of influenza A/H1N1 viruses should be based on needs of laboratories.