Eurofins GeneScan Technologies GmbH, Engesserstraße 4, D-79108 Freiburg im Breisgau, Germany.
Viracor Eurofins, 1001 NW Technology Drive, Lee's Summit, MO 64086, USA.
J AOAC Int. 2021 Aug 20;104(4):872-888. doi: 10.1093/jaoacint/qsab045.
The Eurofins GeneScan Technologies' VIRSeek SARS-CoV-2 Mplex kit is a RT (reverse transcription) real-time polymerase chain reaction (RT-qPCR) assay for the detection of two targets on the N-gene (nucleocapsid) of SARS-CoV-2. An extraction control, that allows monitoring of the extraction procedure and PCR inhibition, is included.
In silico analysis and wet testing showed inclusivity and exclusivity of the assay. The complete workflow starting from surface swabbing (VIRSeek PATHOSwab kit), RNA extraction (VIRSeek RNAExtractor), RT-PCR (VIRSeek SARS-CoV-2 Mplex), and evaluation with FastFinder was validated in comparison to the CDC method for detection of SARS-CoV-2 on stainless steel.
In silico analysis was performed by using the MFOLD online program. The matrix study was performed for stainless steel inoculated with SARS-CoV-2 isolated from the first documented US case of a traveler from Wuhan, China.
For inclusivity, 15 764 sequences were analyzed and all mismatches (0.37% of the sequences had single mismatches) were considered non-critical. Cross reactivity for closely related viruses and background organisms was performed, resulting in correct exclusion of all. No significant differences were observed for the probability of detection (POD) study when comparing to the CDC method.
Results of the inclusivity and exclusivity study show that the assay is specific for detection of SARS-CoV-2. The POD study showed no statistically significant difference compared to the CDC reference method, results were identical for the uninoculated and the high level. For the fractional recovery level, the candidate method detected 9/17 samples leading to a POD of 0.47, the reference method detected 11/20 samples leading to a POD of 0.55.
The complete workflow starting from swabbing of the surface (VIRSeek PATHOSwab kit), RNA extraction (VIRSeek RNAExtractor), RT-PCR (VIRSeek SARS CoV-2 Mplex) and evaluation with FastFinder was validated in comparison to the US Centers for Disease Control and Prevention method for detection of SARS-CoV-2 on Stainless Steel.
Eurofins GeneScan Technologies 的 VIRSeek SARS-CoV-2 Mplex 试剂盒是一种用于检测 SARS-CoV-2 N 基因（核衣壳）上两个靶标的实时 RT-PCR（逆转录）检测试剂盒。试剂盒包含一个提取对照，可用于监测提取过程和 PCR 抑制情况。
通过计算机模拟分析和湿实验检测，评估该检测方法的包容性和排他性。从表面拭子取样（VIRSeek PATHOSwab 试剂盒）、RNA 提取（VIRSeek RNAExtractor）、RT-PCR（VIRSeek SARS-CoV-2 Mplex）到 FastFinder 评估的整个工作流程与美国疾病控制与预防中心（CDC）检测不锈钢表面 SARS-CoV-2 的方法进行了比较验证。
使用在线 MFOLD 程序进行计算机模拟分析。该研究对从首例来自中国武汉旅行者的美国首例确诊病例中分离出的 SARS-CoV-2 接种于不锈钢表面的样本进行了基质研究。
包容性研究共分析了 15764 个序列，所有错配（序列的 0.37%存在单个错配）均被认为是非关键性的。对密切相关的病毒和背景生物体进行了交叉反应性研究，结果均正确排除。与 CDC 方法相比，概率检测（POD）研究未观察到显著差异。
包容性和排他性研究的结果表明，该检测方法可特异性检测 SARS-CoV-2。POD 研究与 CDC 参考方法相比无统计学显著差异，未接种和高接种水平的结果一致。对于部分回收水平，候选方法检测到 17 个样本中的 9 个，导致 POD 为 0.47，参考方法检测到 20 个样本中的 11 个，导致 POD 为 0.55。
与美国疾病控制与预防中心（CDC）检测不锈钢表面 SARS-CoV-2 的方法相比，从表面拭子取样（VIRSeek PATHOSwab 试剂盒）、RNA 提取（VIRSeek RNAExtractor）、RT-PCR（VIRSeek SARS CoV-2 Mplex）和 FastFinder 评估的完整工作流程得到了验证。
