一种快速核酸提取试剂在流感病毒检测中的应用评价

Application of a rapid nucleic acid extraction method for influenza virus detection

目的评价一种快速核酸提取试剂在流感病毒检测中的可应用性,为流感病毒核酸快速提取检测提供技术参考。方法分别使用离心柱法、磁珠法和快速法核酸提取试剂对流感病毒MDCK细胞株的不同稀释度标本以及20份阳性咽拭子标本进行核酸提取,然后进行实时荧光PCR检测,用SPSS 21.0软件对结果进行统计分析,评价核酸快速提取方法的RNA提取效率。结果快速核酸提取试剂对不同稀释度的流感病毒细胞株提取效率要高于离心柱法和磁珠法,Ct值与离心柱法的差值中位数为-1.51(-1.18～-2.01),与磁珠法的差值中位数为-1.09(-0.83～-1.31),咽拭子标本检测结果也显示出快速法核酸提取试剂核酸提取效率高于离心柱法和磁珠法。3种检测方法检出率和重复性差异无统计学意义。结论快速提取试剂具有操作简单、时间短、对设备要求不高等优势,并且核酸提取效率优于离心柱法和磁珠法,提示快速提取试剂可以应用于流感病毒的核酸检测。

Objective To evaluate the effectiveness of a rapid nucleic acid extraction kit for the detection of influenza virus.Methods Different dilutions of influenza viruses(MDCK cell strain)and 20 positive throat swabs for influenza virus were selected in the study,nucleic acids were extracted respectively using spin column method,magnetic beads method and rapid method,then using real-time fluorescent PCR for influenza virus detection.Results were analyzed using software SPSS 21.0.Results For influenza virus from cell culture and throat swabs,the rapid nucleic acid extraction method showed higher extraction efficiency than spin column and magnetic beads method.There were no differences in the detection rate and repeatability among the three methods.Conclusions Rapid nucleic acid extraction method showed advantages such as easy to operate,short time,no cross contamination,lower requirements for equipment and etc.