Flash10即时检测系统在新型冠状病毒核酸检测中的分析性能评价  

Analytical performance evaluation of Flash10 point‑of‑care testing in severe acute respiratory syndrome coronavirus 2 nucleic acid detection

作　　者：韩彦熙 封婉玉 刘聪 陈玉清 张瑞[1] 李金明[1] Han Yanxi;Feng Wanyu;Liu Cong;Chen Yuqing;Zhang Rui;Li Jinming(National Center for Clinical Laboratories,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing Hospital/National Center of Gerontology,Beijing 100730,China;Chinese Academy of Medical Sciences,Peking Union Medical College,Beijing 100730,China)

机构地区：[1]北京医院国家老年医学中心,国家卫生健康委临床检验中心,中国医学科学院老年医学研究院,北京100730 [2]中国医学科学院北京协和医学院,北京100730

出　　处：《中华医学杂志》2024年第47期4323-4329,共7页National Medical Journal of China

基　　金：国家重点研发计划(2022YFC2603800);北京市科技计划(Z221100007922010)。

摘　　要：目的评价Flash10即时检测(POCT)系统在新型冠状病毒(SARS‑CoV‑2)核酸检测中的分析性能。方法采用无生物传染危险性的噬菌体病毒样颗粒(VLPs)样本和灭活的病毒培养液样本,对Flash10全自动核酸检测系统及其配套的SARS‑CoV‑2核酸检测试剂盒进行分析性能评价。通过检测梯度稀释的非变异株、Omicron(BA.5和BF.7)变异株VLPs以及阴性样本,使用Probit回归分析评价该POCT系统的最低检测限(LoD);通过检测3000.00、1000.00、333.33、111.11拷贝/ml浓度的非变异株和Omicron(BA.5和BF.7)变异株VLPs样本,计算各浓度样本ORF1ab基因和N基因循环阈值(Ct)的变异系数(CV),评价该系统的精密度;通过检测可能引起与SARS‑CoV‑2感染症状相似的13种常见呼吸道病原体,评价该系统的分析特异性;通过检测Alpha、Beta、Delta、Omicron等既往全球范围内流行过的变异株VLPs样本,评价该检测系统对SARS‑CoV‑2变异株的检测能力;并通过交叉污染和核酸扩增产物污染风险研究,评价该检测系统的密闭性。结果该POCT系统检测SARS‑CoV‑2非变异株和Omicron(BA.5、BF.7)变异株样本的LoD分别为92.97拷贝/ml(95%CI:63.68~196.27拷贝/ml)、95.49拷贝/ml(95%CI:67.26~200.14拷贝/ml)和99.27拷贝/ml(95%CI:67.77~209.89拷贝/ml),和试剂说明书中宣称的LoD(100拷贝/ml)一致;检测4个浓度水平的SARS‑CoV‑2非变异株和Omicron(BA.5、BF.7)变异株样本时,ORF1ab基因和N基因Ct值的CV分别为2.41%~4.97%和2.29%~4.48%,均<5%;对SARS‑CoV‑2以外的其他13种常见呼吸道病原体进行检测,结果均为阴性;对既往全球范围内流行过的变异株均可正确检出;交叉污染和核酸扩增产物污染风险的评价结果均为阴性。结论Flash10 POCT系统在SARS‑CoV‑2核酸检测中的分析性能良好。Objective To evaluate the analytical performance of Flash10 point‑of‑care testing(POCT)in the detection of severe acute respiratory syndrome coronavirus 2(SARS‑CoV‑2)nucleic acid.Methods The analytical performance evaluation of the Flash10 POCT system and its matching kits was carried out based on the non‑infectious phage virus‑like particles(VLPs)samples and inactivated viral culture samples.The limit of detection(LoD)was evaluated by testing gradient‑diluted non‑variant and Omicron variants(BA.5 and BF.7)VLPs samples and negative samples and using Probit regression analysis.The precision was evaluated by testing non‑variant and Omicron variants(BA.5 and BF.7)VLPs samples with concentrations of 3000.00,1000.00,333.33 and 111.11 copies/ml and calculating coefficient of variation(CV)for cycle threshold(Ct)value of ORF1ab gene and N gene.The analytical specificity was evaluated by detecting 13 common respiratory pathogens that can cause symptoms similar to coronavirus disease 2019(COVID‑19).The testing ability of SARS‑CoV‑2 variants were assessed by detecting the VLPs samples with variants that have been prevalent worldwide(including Alpha,Beta,Delta,Omicron and so on).The sealing of this POCT system was evaluated by studying cross‑contamination and the risk of contamination of nucleic acid amplification products.Results The LoD of the Flash 10 POCT system for detecting non‑variant samples and Omicron variant(BA.5,BF.7)samples were 92.97 copies/ml(95%CI:63.68-196.27 copies/ml)、95.49 copies/ml(95%CI:67.26-200.14 copies/ml)and 99.27 copies/ml(95%CI:67.77-209.89 copies/ml),respectively,which aligned with the LoD(100 copies/ml)claimed by the reagent instructions.When detecting non‑variant and Omicron variant(BA.5,BF.7)samples with four different concentrations,the CV of the Ct value of ORF1ab gene and N gene were 2.41%to 4.97%and 2.29%to 4.48%,respectively,which were all below 5%.The detection results of 13 common respiratory pathogens other than SARS‑CoV‑2 were all negative.The vari

关 键 词：冠状病毒感染 即时检测 分析性能评价 新型冠状病毒 呼吸道病原体 

分 类 号：R511[医药卫生—内科学] R440[医药卫生—临床医学]