Mim Farzana, Reza Md Selim, Rahman Khan Mohammad Jahidur, Karim Nurul, Rahman Mohammad A, Hossain Md Ibrahim, Biswas Rajib
Department of Biochemistry and Molecular Biology, Jahangirnagar University, Dhaka, BGD.
RT-PCR Lab, Bangabandhu Sheikh Mujib Medical College, Faridpur, BGD.
Cureus. 2021 Dec 22;13(12):e20627. doi: 10.7759/cureus.20627. eCollection 2021 Dec.
Background The coronavirus disease 2019 (COVID-19) pandemic has manifested into an unprecedented public health crisis. The rapid spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has facilitated reagent developers to customize and receive authorization for nucleic acid testing kits in a short period, which would have resulted in some shortcomings in the quality parameters of the kits. Consequently, in-house clinical validations of innovative real-time quantitative polymerase chain reaction (RT-qPCR) kits are required. This research aims to determine the sensitivity, specificity, and accuracy of various RT-qPCR kits available in Bangladesh. Methodology A total of 150 samples were obtained from patients with suspected COVID-19 infection when the delta variant was predominant, followed by RNA extraction performed using a nucleic acid isolation kit. Subsequently, three commercially available PCR kits named Sansure (China), STAT-NAT (Sentinel Diagnostics, Italy), and Roche Biochem (Switzerland) were applied to detect SARS-CoV-2. Results The results showed that the STAT-NAT kit is more sensitive than the other two, as indicated by the cycle threshold (Ct) values of respective genes. STAT-NAT RT-qPCR can detect the gene sensitively (p < 0.001) compared to Sansure. STAT-NAT was also capable of detecting and genes more sensitively (p < 0.001) compared to Roche. Regarding specificity, STAT-NAT (95% confidence interval [Cl] = 92.29-99.73%). RT-qPCR showed more accuracy than Sansure (95% Cl = 90.77-99.32%) and Roche (95% Cl = 81.17-94.38%). The area under the curve for , , and genes of the STAT NAT PCR kit was 0.952, 0.959, and 0.981, respectively. Conclusions This study concluded that STAT-NAT is a better diagnostic RT-qPCR kit compared to Sansure and Roche for detecting SARS-CoV-2.
背景 2019 冠状病毒病（COVID-19）大流行已演变成一场前所未有的公共卫生危机。严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）的迅速传播促使试剂开发商在短时间内定制并获得核酸检测试剂盒的授权，这可能导致试剂盒的质量参数存在一些缺陷。因此，需要对创新的实时定量聚合酶链反应（RT-qPCR）试剂盒进行内部临床验证。本研究旨在确定孟加拉国现有的各种 RT-qPCR 试剂盒的灵敏度、特异性和准确性。方法 当德尔塔变异株占主导时，从疑似 COVID-19 感染患者中总共采集了 150 份样本，随后使用核酸提取试剂盒进行 RNA 提取。随后，应用三种市售的 PCR 试剂盒，分别是深圳华大基因股份有限公司（中国）的Sansure、哨兵诊断公司（意大利）的STAT-NAT和罗氏诊断（瑞士）的Roche Biochem，来检测 SARS-CoV-2。结果 结果表明，根据各基因的循环阈值（Ct）值，STAT-NAT 试剂盒比其他两种试剂盒更灵敏。与 Sansure 相比，STAT-NAT RT-qPCR 能更灵敏地检测该基因（p < 0.001）。与罗氏相比，STAT-NAT 也能更灵敏地检测该基因和该基因（p < 0.001）。在特异性方面，STAT-NAT（95%置信区间[Cl]=92.29-99.73%）。RT-qPCR 显示出比 Sansure（95% Cl = 90.77-99.32%）和罗氏（95% Cl = 81.17-94.38%）更高的准确性。STAT NAT PCR 试剂盒的该基因、该基因和该基因的曲线下面积分别为 0.952、0.959 和 0.981。结论 本研究得出结论，在检测 SARS-CoV-2 方面，与 Sansure 和罗氏相比，STAT-NAT 是一种更好的诊断 RT-qPCR 试剂盒。
