新型冠状病毒大规模核酸筛检扩增试剂保存方式研究

Research on the Preservation Method of Large-scale Nucleic Acid Screening Amplification Reagent of COVID-19

为了研究新型冠状病毒核酸扩增试剂解冻后不同保存方式是否影响核酸检测结果,将新型冠状病毒荧光定量PCR检测试剂盒解冻后采取室温保存、4℃冷藏保存、-20℃冷冻保存2 h、4 h、6 h、12 h、24 h、72 h后分别与高浓度阳性样本和弱阳性样本进行核酸扩增,记录各样本扩增结果及循环数(Ct值)。研究结果表明所有样本均检出阳性结果,扩增试剂3种保存方式下弱阳性样本N基因、ORF1ab基因Ct值不同,差异无统计学意义(P>0.05)。研究初步证实了新型冠状病毒核酸扩增试剂解冻后72 h内采用室温、冷藏、冷冻3种保存方式不会影响高浓度阳性样本和弱阳性样本检出。在大规模人群筛检时,剩余核酸扩增试剂在稳妥保存不受污染情况下仍可用于后续实验。

In order to explore whether the different storage methods of SARS-CoV-2 nucleic acid amplification agents after thawing affect the results of SARS-CoV-2 nucleic acid detection.SARS-CoV-2 fluorescence quantitative PCR detection kit after thawing was stored at room temperature,refrigerated at 4℃,and freezed at-20℃for 2 h,4 h,6 h,12 h,24 h and 72 h.Then,kit was used for nucleic acid amplification with high concentration positive samples and weak positive samples,and the amplification results and cycle threshold value(Ct value)of each sample were recorded.The results show that positive results were detected in all samples,and the Ct values of N gene and ORF1ab gene in the weak positive samples were different under the three storage methods of amplification reagent,and the difference was not statistically significant(P>0.05).This study preliminarily confirmed that the three storage methods of room temperature,refrigeration,and freezing within 72 h after thawing of SARS-CoV-2 nucleic acid amplification reagent would not affect the detection of high concentration positive samples and weak positive samples.Therefore,when screening large-scale populations,the remaining nucleic acid amplification reagents could be used for subsequent experiments when they are stored safely without contamination.