4种核酸提取方法在流行性感冒病毒核酸检测中的比较

Comparison of four RNA extraction methods for detection of influenza viruses nucleic acids

目的：比较常见的4种核酸抽提方法，比较其对流行性感冒（流感）病毒检测的敏感性差异，为实验室应用和结果的评价提供依据。方法：对已确定效价的甲1型流感病毒，作10^-2～10^-5系列稀释，选用Roche公司Hish Pure Viral RNA Kit、QIAGEN公司的Rneasy mini Kit、国产Z生物公司RNA提取试剂盒和Invitrogen公司Trizol试剂等4种核酸提取方法提取上述标本核酸模板，采用罗氏公司的RT-PCR试剂盒，用实时荧光定量聚合酶链反应（FQ-PCR）对其进行评价。结果：在对不同病毒拷贝数流感病毒的检测中，4种核酸提取方法以Roche公司High Pure Viral RNA Kit提取效率最高，以此提取效率作为100％，则QIAGEN Rneasy mini Kit、Invitrogen Trizol和国产Z生物公司RNA提取试剂盒的平均提取效率依次为89．71％、75．30％、68．00％。经方差分析及t检验，4种方法之间存在显著性差异（P均〈0．05）。对临床标本的检测，也获得相似结果。结论：各试剂之间提取核酸的效率存在差异，应该选择性价比较好的方法进行临床标本的实验室RNA提取。

Objective： To compare the efficiency of four RNA extraction methods for the detection of influenza （flu） viruses. Methods： We compared the efficiency of four RNA extraction methods including High Pure Viral RNA Kit （ Roche ） , QIAamp Rneasy mini Kit（Qiagen） ,RNA extraction Kit（Z Bioteeh Co. Ltd in China） and Trizol（Invitrogen）. The specimens tested were the 10^-2 - 10^-5 successive dilution of a known flu viruses and 5 throat washing samples collected from the patients with acute respiratory tract infection nucleic acid of flu viruses RNA were extracted using each RNA extraction methods and detected by fluorescence real- time quantitative RT- PCR（Taqman）. Results：The extraction efieieney of four RNA extraction methods had significant statistical difference （P 〈0. 05） in the detection of the dilution flu vimsos. The extraction efieieney of the High Pure Viral RNA Kit （Roche） extraction method was the highest （relative 100% ） and the QIAamp Rneasy mini Kit（Qiagen）, Trizol （Invitrogen） and RNA extraction Kit （Z biological Co, Ltd in China） extraction methods were lower （relative 89. 71% ,75. 30%, 68.00% respectively ）. To clinical specimens, the High Pure Viral RNA Kit （Roche） extraction method was also the best. Conclusion：The results suggest that it had significant difference between four RNA extraction methods, so we should choose the appropriate one for detection of flu viruses in clinical specimens.