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 3 000.00, 1 000.00, 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 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. Conclusion: The Flash10 POCT system demonstrates good analytical performance for SARS-CoV-2 nucleic acid detection.
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3 000.00、1 000.00、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。.