ABSTRACT

Objectives

Sensitive molecular diagnostic assays are essential for COVID-19 diagnosis. We evaluated the Hecin Scientific SARS-CoV-2 nucleic acid test kit, a dual-target real-time RT-PCR assay targeting the SARS-CoV-2 N and ORF1ab genes.

Methods

The Hecin test kit’s diagnostic performance in detecting SARS-CoV-2 RNA was compared to the LightMix Modular SARS and Wuhan CoV E-gene kit (TIB Molbiol) and an in-house single-tube nested real-time RT-PCR using 296 clinical specimens, 11 proficiency testing samples, and 30 low-positive deep throat saliva and nasopharyngeal swab (NPS) samples pooled into negative samples in ratios of 1:5, 1:10, and 1:30.

Results

The limit-of-detection of the Hecin test kit was around 500 dC/mL for the N and ORF1ab targets. Sensitivity and specificity of the Hecin test kit were 98.1% (95% CI: 93.4–99.8%) and 100% (98.1–100%), respectively, when measured against the reference method. The Hecin test kit showed fair sensitivity (80%) in low-positive NPS samples pooled in ratios of 1:5 and 1:10. Its performance in pooled samples could be dramatically improved by adjusting the assay Ct cutoff.

Conclusion

The Hecin test kit enables sensitive and specific detection of SARS-CoV-2 in clinical samples and pooled samples.

中文翻译：

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用于 COVID-19 诊断的双靶标实时 RT-PCR 检测的比较评估和混合唾液和鼻咽拭子样本的性能评估

摘要

目标

敏感的分子诊断分析对于 COVID-19 诊断至关重要。我们评估了 Hecin Scientific SARS-CoV-2 核酸检测试剂盒，这是一种针对 SARS-CoV-2 N 和 ORF1ab 基因的双靶标实时 RT-PCR 检测。

方法

Hecin 检测试剂盒在检测 SARS-CoV-2 RNA 方面的诊断性能与 LightMix Modular SARS 和武汉冠状病毒 E 基因试剂盒 (TIB Molbiol) 以及使用 296 个临床样本的内部单管嵌套实时 RT-PCR 进行了比较。样本、11份能力验证样本和30份低阳性深喉唾液和鼻咽拭子（NPS）样本按1:5、1:10和1:30的比例合并为阴性样本。

结果

对于 N 和 ORF1ab 目标，Hecin 检测试剂盒的检测限约为 500 dC/mL。与参考方法相比，Hecin 检测试剂盒的灵敏度和特异性分别为 98.1% (95% CI: 93.4–99.8%) 和 100% (98.1–100%)。Hecin 检测试剂盒在以 1:5 和 1:10 的比例混合的低阳性 NPS 样本中显示出相当的灵敏度 (80%)。通过调整检测 Ct 截止值，可以显着提高其在混合样品中的性能。

结论

Hecin 检测试剂盒能够灵敏、特异地检测临床样本和混合样本中的 SARS-CoV-2。