摘要
目的比较磁珠吸附法和样本释放剂法对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核酸检测结果的符合率、重复性及检出限,优选出提取效率高、耗时短、成本低的核酸提取方法。方法采用2种核酸提取方法的3种商品化核酸提取试剂,分别对阴性样本、阳性样本及不同浓度(稀释度20~28)的SARSCoV-2 RNA标准品进行RNA提取及实时荧光逆转录聚合酶链反应(RT-PCR)检测,以Ct值评价不同核酸提取方法的性能。结果3种核酸提取试剂的阴性和阳性样本结果符合率均为100%。使用磁珠吸附法进行核酸提取的变异系数(CV)(CV为0.89%~1.58%)低于样本释放剂法,样本释放剂B的CV(CV为0.83%~2.73%)次之,样本释放剂A的CV最大(CV为1.62%~4.90%)。磁珠吸附法及样本释放剂B提取核酸的检出限均为156.3拷贝/mL,样本释放剂A的检出限为625.0拷贝/mL。结论磁珠吸附法核酸提取试剂核酸提取效率高、重复性好,适合大批量样本的集中操作。
Objective To compare the consistency rate,repeatability and detection limit of magnetic bead adsorption method and sample releasing agent method in the nucleic acid extraction of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),so as to optimize the extraction method of nucleic acid with high extraction efficiency,short time consuming and low cost.Methods Totally,3 commercial nucleic acid extraction reagents for 2 methods of nucleic acid extraction were used to extract RNA from negative samples,positive samples and SARS-CoV-2 standard samples with different dilution(20-28),and real-time fluorescence reverse transcription-polymerase chain reaction(RT-PCR)was performed.Ct value was used to evaluate the performance of different nucleic acid extraction methods.Results The consistency rate of negative and positive samples of 3 nucleic acid extraction reagents was 100%.The coefficient of variation(CV)of magnetic bead adsorption method was lower than those of sample releasing agent method(CV 0.89%-1.58%),sample releasing agent A(CV 1.62%-4.90%)and sample releasing agent B(CV 0.83%-2.73%).The detection limit of magnetic bead adsorption method and sample releasing agent B was 156.3 copies/mL,and that of sample releasing agent A was 625.0 copies/mL.Conclusions The magnetic bead adsorption method of nucleic acid extraction is efficient and reproducible,which is suitable for the centralized operation of a large number of samples.
作者
张云丽
王鑫
邵玲
曲波
赵鸿梅
ZHANG Yunli;WANG Xin;SHAO Ling;QU Bo;ZHAO Hongmei(Department of Clinical Laboratory,Liaoning Provincial People's Hospital,Shenyang 110016,Liaoning,China)
出处
《检验医学》
CAS
2021年第5期530-534,共5页
Laboratory Medicine
关键词
严重急性呼吸综合征冠状病毒2
核酸检测
核酸提取
实时逆转录荧光聚合酶链反应
Severe acute respiratory syndrome coronavirus 2
Nucleic acid detection
Nucleic acid extraction
Real-time fluorescence reverse transcription-polymerase chain reaction
