摘要
目的选择常见的3种核酸提取方法,比较其对低拷贝数甲型H1N1流感病毒核酸检测的敏感性差异,为实验室应用和结果的评价提供依据。方法对已确定阳性的甲型H1N1流感病毒标本,作10-1～10-4系列稀释,选用Promega公司全自动核酸提取仪及配套试剂盒、QIAGEN公司Rneasy MiniKit和国产H公司提取试剂盒3种核酸提取方法提取上述标本核酸模板,采用实时荧光定量逆转录聚合酶链反应(荧光RT-PCR)对其进行评价。结果在对不同拷贝数流感病毒的检测中,3种核酸提取方法以Promega公司全自动核酸提取仪提取效率最高,以此提取效率作为100%,则QIAGEN Rneasy MiniKit和国产H公司提取试剂盒的平均提取效率依次为83.27%、55.70%。经方差分析,3种方法之间存在显著性差异(P<0.05)。结论各试剂之间提取核酸的效率存在显著差异,建议根据实际情况采用合适的核酸提取试剂和方法。
Aim To compare the efficiency of three RNA extraction methods for detection of HIN1 influenza （flu） viruses. Methods The efficiency of three RNA extraction methods including automated nucleic acid extraction method （Promega）,Rneasy Mini Kit （Qiagen）,RNA extraction Kit （H Biotech Co.Ltd in China） were compared and the specimens tested were the 10^-1 -10^-4 serial dilution of twenty known H1N1 flu viruses throat washing samples collected from the patients with a cute 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 Kit （Daangene）. Results The extraction eficiency of three RNA extraction methods＇ had significant statistical difference （P〈0.05） in the detection of the dilution flu viruses. The extraction efficiency of the automated total nucleic acid extraction method （Promega） was the highest （relative 100%） and the Rneasy mini Kit （Qiagen）,and RNA extraction Kit （ H biological Co.Ltd in China） extraction methods were lower （relative 83.27%、55.70% ,respectively）. Conclusion The results indicate that it had significant difference between three RNA extraction methods, suggesting that the appropriate one should be choose for detection of flu viruses in clinical specimens.
出处
《中国热带医学》
CAS
2010年第7期783-785,共3页
China Tropical Medicine
关键词
甲型N1N1病毒
全自动核酸提取
提取效率
实时荧光定量逆转录聚合酶链反应
H1N1 influenza virus
Automated nucleic acid extraction
Efficiency of extraction
Real-time fluorescence quantitative reverse transcription polymerase chain reaction
