新型冠状病毒混合样品检测研究

doi:10.6040/j.issn.1671-7554.0.2021.0126

摘要/Abstract

摘要： 目的探究在新型冠状病毒样本混合检测中,稀释混样检测和混采检测的合理样本数量,为大人群样本的核酸筛查提供更为快捷有效的检测方案。方法设计一种利用注射器纯化柱进行大体积样本保存液核酸提取方法,在稀释混样检测和混采检测中,对比使用该方法将全部样本保存液提取核酸和只取200 μL样本保存液提取核酸时检测结果的差异。结果稀释混样检测中,对于单独一份阳性样本,取全部保存液样本提取核酸时ORF1ab基因和N基因的Ct值比取200 μL保存液样本提取核酸时分别小3.583、2.904;当1份阳性样本与9份、19份阴性样本混合后,将全部样本保存液提取核酸,不同混合人份检测的Ct值差异无统计学意义(P>0.05);当混合样本多于20份时,Ct值明显变大。混采检测中,取所有保存液样本提取核酸检测所得Ct值明显小于只取200 μL保存液样本提取核酸所得Ct值(目前通行方案)。结论在新型冠状病毒混合检测中,采用注射器纯化柱法将全部样本的混合保存液提取核酸,稀释混样检测时,可将混合人数上限由20人提高到50人;采用混采检测时,可以明显降低核酸检测的Ct值,从而提高检出率,减少混合检测过程中假阴性现象的发生。

关键词: 新型冠状病毒, 稀释混样检测, 混采检测, 注射器纯化柱, RT-PCR

Abstract: Objective To determine the reasonable sample sizes of diluted mixed sample test and n-in-1 test in the detection of SARS-CoV-2, so as to provide a more efficient scheme for nucleic acid screening of large samples. Methods A method was designed to extract nucleic acid from the preservation solution of large volume samples using a syringe purification column. In both diluted mixed sample test and n-in-1 test, this method was used in the extraction of nucleic acid from the total preservation solution and from 200 μL solution. The test results were compared. Results In diluted mixed sample test, for a single positive sample, the Ct values of ORF1ab gene and N gene were 3.583 and 2.904 lower from the total preservation solution than from the 200 μL solution; when one positive sample was mixed with 9 negative samples and 19 negative samples, there was no statistical difference in Ct values between the two sample sizes(P>0.05); when the number of mixed samples was more than 20, the Ct values increased significantly. In the n-in-1 test, the Ct values of nucleic acid extracted from total preservation solution was significantly lower than that from 200 μL solution(the current scheme). Conclusion In mixed sample detection, using a syringe purification column to extract nucleic acid from the total preservation solution can increase the maximum number of sample size from 20 to 50. With this strategy, the Ct values of nucleic acid test can be significantly reduced, so as to improve the detection rate and reduce the number of false-negative cases in n-in-1 test.

Key words: Severe Acute Respiratory Syndrome Coronavirus 2, Diluted mixed sample test, n-in-1 test, Syringe purification column, RT-PCR

中图分类号:

R574

董宏杰,张俊梅,王帅,王宏伟,张坤迪,胡玮,谢晓鸿,谢时灵,谷立川. 新型冠状病毒混合样品检测研究[J]. 山东大学学报 (医学版), 2021, 59(4): 1-5.

DONG Hongjie, ZHANG Junmei, WANG Shuai, WANG Hongwei, ZHANG KundiHU Wei, XIE Xiaohong, XIE Shiling, GU Lichuan. Research on mixed sample test for SARS-CoV-2[J]. Journal of Shandong University (Health Sciences), 2021, 59(4): 1-5.

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