Han Y X, Feng W Y, Liu C, Chen Y Q, Zhang R, Li J M
National Center for Clinical Laboratories, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital/National Center of Gerontology, Beijing100730, China.
Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing100730, China.
Zhonghua Yi Xue Za Zhi. 2024 Dec 17;104(47):4323-4329. doi: 10.3760/cma.j.cn112137-20240719-01661.
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. 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 3 000.00, 1 000.00, 333.33 and 111.11 copies/ml and calculating coefficient of variation () 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. 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%: 63.68-196.27 copies/ml)、95.49 copies/ml (95%: 67.26-200.14 copies/ml) and 99.27 copies/ml (95%: 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 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 variants that have been prevalent worldwide can be correctly detected. The evaluation results of the risk of cross-contamination and nucleic acid amplification products contamination indicated that the Flash 10 POCT testing system was well sealed. The Flash10 POCT system demonstrates good analytical performance for SARS-CoV-2 nucleic acid detection.
评估Flash10即时检测（POCT）在检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）核酸方面的分析性能。基于非感染性噬菌体病毒样颗粒（VLPs）样本和灭活病毒培养样本，对Flash10 POCT系统及其配套试剂盒进行分析性能评估。通过检测梯度稀释的非变异株和奥密克戎变异株（BA.5和BF.7）VLPs样本及阴性样本，并采用概率回归分析来评估检测限（LoD）。通过检测浓度为3000.00、1000.00、333.33和111.11拷贝/毫升的非变异株和奥密克戎变异株（BA.5和BF.7）VLPs样本，并计算开放阅读框1ab（ORF1ab）基因和N基因循环阈值（Ct）值的变异系数（）来评估精密度。通过检测13种可引起与2019冠状病毒病（COVID-19）相似症状的常见呼吸道病原体来评估分析特异性。通过检测含有全球流行变异株（包括阿尔法、贝塔、德尔塔、奥密克戎等）的VLPs样本，评估SARS-CoV-2变异株的检测能力。通过研究交叉污染和核酸扩增产物污染风险来评估该POCT系统的密封性。Flash 10 POCT系统检测非变异株样本和奥密克戎变异株（BA.5、BF.7）样本的LoD分别为92.97拷贝/毫升（95%：63.68 - 196.27拷贝/毫升）、95.49拷贝/毫升（95%：67.26 - 200.14拷贝/毫升）和99.27拷贝/毫升（95%：67.77 - 209.89拷贝/毫升），与试剂说明书声称的LoD（100拷贝/毫升）相符。在检测四种不同浓度的非变异株和奥密克戎变异株（BA.5、BF.7）样本时，ORF1ab基因和N基因Ct值的变异系数分别为2.41%至4.97%和2.29%至4.48%，均低于5%。13种除SARS-CoV-2外的常见呼吸道病原体检测结果均为阴性。可正确检测全球流行的变异株。交叉污染和核酸扩增产物污染风险的评估结果表明，Flash 10 POCT检测系统密封性良好。Flash10 POCT系统在SARS-CoV-2核酸检测方面表现出良好的分析性能。
