磁珠法提取新型冠状病毒核酸的稳定性  

Stability of SARS-CoV-2 Nucleic Acid Extracted by Magnetic Bead Method

作　　者：秦冬梅[1] 杨志[1] 肖光军[2] 周波[1] 曾玉兰 Qin Dongmei;Yang Zhi;Xiao Guangjun;Zhou Bo;Zeng Yulan(Daying County People's Hospital,Daying Sichuan 629300,China;Suining Central Hospital,Suining Sichuan 629000,China)

机构地区：[1]大英县人民医院,四川大英629300 [2]遂宁市中心医院,四川遂宁629000

出　　处：《医疗装备》2023年第12期9-12,17,共5页Medical Equipment

摘　　要：目的探索磁珠法提取新型冠状病毒(SARS-CoV-2)核酸及其与试剂混合后的稳定性。方法将国家卫生健康委员会临床检验中心(NCCL)和四川省临床检验中心(SPCCL)提供的32份SARS-CoV-2室间质量评价(EQA)样本分为4组,每组含3份NCCL样本和5份SPCCL样本。所有样本均采用磁珠法提取核酸,得到的核酸采用实时荧光逆转录聚合酶链反应(RT-PCR)法立即检测1次(靶基因为ORF1ab基因和N基因),即为各样本的初始结果。各样本余下的核酸按以下方式处理:第1组和第2组样本的核酸分别于4℃和20℃保存,在第6、12、18、24天各检测1次并记录结果;第3组和第4组样本的核酸与扩增试剂混合组成反应体系,再分别于4℃和20℃保存,在第4、8、16小时各检测1次并记录结果。所有结果用循环阈值(即Ct值)表示。核酸保存后的各次检测结果与其初始结果进行比较。结果磁珠法提取SARS-CoV-2核酸于4℃和20℃保存24 d内各次检测结果与初始结果比较,差异均无统计学意义(P>0.05);磁珠法提取SARS-CoV-2核酸与扩增试剂混合后,于4℃和20℃保存16 h内各次检测结果与初始结果比较,差异均无统计学意义(P>0.05)。结论4℃或20℃保存条件下,磁珠法提取SARS-CoV-2核酸至少可以稳定24 d,与试剂混合后至少可以稳定16 h。Objective The severe acute respiratory syndrome coronavirus 2(SARSCoV-2)nucleic acid extracted by magnetic bead method and its stability after mixing with reagent were explored.Methods 32 EQA samples of SARS-CoV-2 provided by the National Center for Clinical Laboratories(NCCL)and Sichuan Province Center for Clinical Laboratories(SPCCL)were divided into 4 groups,with each group containing 3 NCCL samples and 5 SPCCL samples.With nucleic acid extracted with magnetic bead method in all samples,the obtained nucleic acid was immediately detected once using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR)method(ORF1ab and N genes as the target genes),which was the initial result of each sample.The remaining nucleic acid of each sample was processed as follows:the nucleic acid of group 1 and group 2 samples were stored at 4℃and 20℃,respectively,and tested once on the 6th,12th,18th,and 24th day,and the results were recorded;The nucleic acid of group 3 and group 4 samples were mixed with amplification reagents to form the reaction system,and then stored at 4℃and 20℃,respectively.The test was performed once at 4,8 and 16 hours,and the results were recorded.With the all results represented by cycle thresholds(Ct values),the results of each test after nucleic acid preservation were compared with the initial results.Results The SARS-CoV-2 nucleic acid extracted by the magnetic bead methodwas stored at 4℃and 20℃for 24 days,and there were no statistically significant differencesbetween the test results and the initial results(P>0.05);After mixing SARS-CoV-2 nucleicacid extracted by magnetic bead method with amplification reagent,there was no statisticalsignificance between the results of each test and the initial results within 16 h stored at 4℃and20℃(P>0.05).Conclusion SARS-CoV-2 nucleic acid extracted by magnetic beads methodcan be stable for at least 24 days at 4℃or 20℃,and can be stable for at least 16 hours aftermixing with reagents.

关 键 词：新型冠状病毒 核酸提取 磁珠法 稳定性 

分 类 号：R373.1[医药卫生—病原生物学]