SARS-CoV-2核酸检测弱阳性质控品的自制及效果评价  

Evaluation of self preparation and efficacy of weakly positive quality control product for SARS-CoV-2 nucleic acid detection

作　　者：祝惠钦 梁瑞健 谢荣章 陈树华 何小霞 彭世姚 ZHU Hui-qing;LIANG Rui-jian;XIE Rong-zhang(YunFu People's Hospital,GuangDong Yunfu 527300)

机构地区：[1]云浮市人民医院检验科,广东云浮527300

出　　处：《医学检验与临床》2025年第2期17-24,共8页Medical Laboratory Science and Clinics

基　　金：云浮市医学科学技术研究项目,项目编号:2022B010。

摘　　要：目的:本研究旨在通过实验室内稀释法自配新型冠状病毒弱阳性质控品,并对其性能进行评价以满足新冠病毒核酸检测的质量控制需求。方法:使用阴性临床样本、样本保存液和去离子水三种基质将强阳性假病毒核酸质控品稀释成弱阳性质控品,采用逆转录实时荧光定量PCR(RT-qPCR)进行核酸扩增,评价其分装后的均一性以及其在反复冻融、室温放置、2~8℃、-20℃、-70℃的不同条件下的稳定性。结果:三种基质配制的质控品分装后均一性良好,使用阴性临床样本制备的弱阳性质控品能在反复冻融4次、室温放置20小时、2~8℃暂存6天、-20℃冻存8周和-70℃冻存16周保持稳定;而使用样本保存液配制的弱阳性质控品能在室温放置16小时、2~8℃暂存6天、-20℃冻存6周和-70℃冻存20周保持稳定;去离子水组则在冻融1次不能稳定。结论:阴性临床样本和保存液均是实验室内稀释法自配新型冠状病毒弱阳性质控品理想的基质,并且稳定性能满足临床日常检测需求。建议根据实验室实际条件选择,并结合所使用扩增试剂检测靶基因覆盖范围,确定最佳的使用条件,保证检测质量。Objective:The objective of this study was to create a weakly positive quality control product for the new coronavirus through in-laboratory dilution and to assess its effectiveness in meeting the quality control requirements of new coronavirus nucleic acid detection assays.Methods:The strong-positive pseudovirus nucleic acid quality control was diluted into a weak-positive quality control using three matrices:negative clinical samples,sample preservation solution and deionised water,and nucleic acid amplification was performed by reverse transcription real-time fluorescence quantitative PCR(RT-qPCR)to evaluate the homogeneity of the dispensed sample as well as its stability under different conditions,including repeated freezing and thawing,placing at room temperature,2-8℃,-20℃,and-7oC.Results:The homogeneity of the quality controls prepared from the three matrices was good after dispensing,The weakpositive quality controls prepared from negative clinical samples remained stable after four freeze-thaw cycles,20 h at room temperature,6 days of intermediate storage at 2-8℃,8 weeks of freezing at-20℃ and 16 weeks of freezing at-70℃;The weak-positive quality controls prepared from sample preservation solutions remained stableafter 16 h atroom temperature 6 days ofstorage at 2-8℃,6 weeks offreezing at-20℃ and 20 weeks of freezing at-70℃,However,Deionised Water group can not be stabilized during one freeze-thaw.Conclusion:Negative clinical samples and sample preservation solutions are suitable matrices for self-assembly of novel coronavirus weak-positive quality controls using in-laboratory dilution methods.These matrices are stable enough to meet routine clinical testing requirements It is recommended that the choice ofreagent be made according to the actual conditions ofthe laboratory.In conjunction with the coverage of target genes detectedby the amplification reagents used,the optimal conditions ofuse are determined in order to ensure the quality ofthe test.

关 键 词：新型冠状病毒 弱阳性核酸质控品 均一性 稳定性 

分 类 号：R44[医药卫生—诊断学]