摘要
目的评估A、B、C、D 4种新型冠状病毒(2019-nCoV)核酸检测试剂盒的分析性能,以便选择性能佳、性价比高的试剂用于临床检测。方法①将国家卫生健康委临床检验中心和湖北省临床检验中心2021年2019-nCoV(含变异株)核酸检测室间质量评价样本36例(27例阳性和9例阴性),磁珠法提取核酸后分别应用4种试剂扩增以评价其符合率;②应用2019-nCoV阴性混合鼻咽拭子样本将商品化2019-nCoV RNA液体室内质控品(以ORF1ab浓度赋值17300 copies/mL)稀释12倍到1441.7 copies/mL,连续5 d每天做5个复孔,共计25孔,磁珠法提取核酸后分别应用4种试剂扩增以评价其重复性和实验室内精密度;③应用2019-nCoV阴性混合鼻咽拭子样本将2019-nCoV RNA液体室内质控品(以ORF1ab浓度赋值17300 copies/mL)梯度稀释成567.7、216.3和108.2 copies/mL 3种浓度,连续3 d每种浓度每天做7~8个复孔,共计23个复孔,磁珠法提取核酸后分别应用4种试剂扩增以评价其分析灵敏度。结果①对36例2019-nCoV核酸检测室间质量评价(EQA)样本,试剂A、B和D的符合率均为100%(36/36),试剂C的符合率为97.2%(35/36),但试剂A、B和C分别有2、6和10个样本需要复查后才能判定结果。②4种试剂检测1441.7 copies/mL 2019-nCoV RNA质控品稀释液的靶基因循环阈值(Ct)重复性和实验室内精密度均小于5%。③4种试剂A、B、C、D对576.7 copies/mL 2019-nCoV RNA质控品稀释液的检出率分别为95.7%、34.8%、8.7%和100%,对216.3 copies/mL稀释液的检出率则分别为95.7%、13.0%、17.4%和91.3%,A和D的分析灵敏度明显优于B和C。结论4种新型冠状病毒核酸检测试剂的分析性能存在差异,D最优,A次之,明显优于B和C。
Objective To assess the analytic performance of four different 2019-nCoV nucleic acid detection kits(labeled A,B,C and D respectively),and select the kits with good analytic performance and high cost performance for clinical laboratory.Methods①These 36 samples(27 positive and 9 negative for 2019-nCoV RNA)for external quality assessment(EQA)from National Center For Clinical Laboratories(NCCL)and Hubei Center For Clinical Laboratory(HBCCL)were extracted and tested using these four kits to evaluate overall agreement.②The 2019-nCoV pseudovirus with 17300 copies/mL of ORF1ab measured by droplet digital PCR was diluted with clinical pooled 2019-nCoV negative nasopharyngeal swab specimens to 1441.7 copies/mL.The 1441.7 copies/mL of 2019-nCoV pseudovirus was extracted and tested five times per day in a single run for 5 consecutive days using these four kits,and the repeatability and within-laboratory precision were calculated following EP15-A3.③The 567.7,216.3 and 108.2 copies/mL of 2019-nCoV pseudovirus were extracted and tested seven or eight times per day in a single run for 3 consecutive days using these four kits.The positive rate of each dilution was counted and the analytic sensitivity was evaluated.Results①For these 36 EQA samples,the concordance rates were 100%(36/36)for A,B and D,and 97.2%(35/36)for C,respectively.But there were 2,6,and 10 EQA specimens which needed to be tested again by kit A,B and C respectively.②The repeatability and within-laboratory precision(%CV)of the cycles threshold(Ct)of the target genes tested by these four kits at 1441.7 copies/mL were less than 5%.③The detection rates of 576.7 copies/mL of 2019-nCoV pseudovirus were 95.7%for A,34.8%for B,8.7%for C and 100%for D,respectively.The detection rates of 216.3 copies/mL of 2019-nCoV pseudovirus were 95.7%for A,13.0%for B,17.4%for C and 91.3%for D,respectively.Conclusion The analytic performance of these four 2019-nCoV RNA detection kits was different.The analytic performance of D was the best,followed by A,and the analytic perf
作者
陈家颖
黄达
耿帜
陈凤花
CHEN Jiaying;HUANG Da;GENG Zhi;CHEN Fenghua(Department of Clinical Laboratory,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,China)
出处
《临床血液学杂志》
CAS
2022年第8期547-551,共5页
Journal of Clinical Hematology
关键词
新型冠状病毒
精密度
分析灵敏度
实时荧光PCR
2019-nCoV
precision
analytic sensitivity
real-time fluorescent polymerase chain reaction
