Visseaux Benoit, Collin Gilles, Houhou-Fidouh Nadhira, Le Hingrat Quentin, Ferré Valentine Marie, Damond Florence, Ichou Houria, Descamps Diane, Charpentier Charlotte
Université de Paris, INSERM UMR 1137 IAME, Laboratoire de Virologie, AP-HP, Hôpital Bichat-Claude Bernard, F-75018 Paris, France.
Université de Paris, INSERM UMR 1137 IAME, Laboratoire de Virologie, AP-HP, Hôpital Bichat-Claude Bernard, F-75018 Paris, France.
J Virol Methods. 2021 May;291:114086. doi: 10.1016/j.jviromet.2021.114086. Epub 2021 Feb 9.
The worldwide demand for SARS-CoV-2 RT-PCR testing resulted in a shortage of diagnostic kits. RNA extraction step constitutes a major bottleneck to perform diagnostic. The aim of this study was to assess performances of different extraction-free SARS-CoV-2 RT-PCR assays compared to a reference RT-PCR assay. The panel of evaluation consisted of 94 samples: 69 positive and 25 negative for SARS-CoV-2 by reference RT-PCR. Three extraction-free RT-PCR assays were assessed: (i) PrimeDirect® Probe RT-qPCR Mix (Takara), (ii) PrimeScript®RT-PCR (Takara), and (iii) SARS-CoV-2 SANSURE®BIOTECH Novel Coronavirus (Sansure). The overall sensitivity of PrimeDirect, PrimeScript and Sansure assays was 55.1 %, 69.6 % and 69.6 %, respectively. The sensitivity increased among samples with Ct<30: 91.9 % (n = 34/37), 89.2 % (n = 33/37) and 94.6 % (n = 35/37) for PrimeDirect, PrimeScript and Sansure assays, respectively. The specificity was 88 %, 100 % and 100 % for PrimeDirect, PrimeScript and Sansure assays, respectively. In the present study, we showed a good sensitivity of extraction-free PCR assays, especially for high viral loads (Ct<30), except PrimeDirect that displayed imperfect sensitivity and specificity. Despite a lower sensitivity for low viral loads, extraction-free reagents can provide a valuable option, cheaper, easier and less reagent consuming for SARS-CoV-2 diagnostic, especially in laboratory with lower experience and equipment for molecular assays.
全球对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）逆转录聚合酶链反应（RT-PCR）检测的需求导致诊断试剂盒短缺。RNA提取步骤是进行诊断的主要瓶颈。本研究的目的是评估不同的免提取SARS-CoV-2 RT-PCR检测方法与参考RT-PCR检测方法相比的性能。评估样本组由94个样本组成：通过参考RT-PCR检测，69个样本为SARS-CoV-2阳性，25个样本为阴性。评估了三种免提取RT-PCR检测方法：（i）PrimeDirect®探针RT-qPCR Mix（Takara），（ii）PrimeScript®RT-PCR（Takara），以及（iii）SARS-CoV-2 SANSURE®BIOTECH新型冠状病毒（Sansure）。PrimeDirect、PrimeScript和Sansure检测方法的总体灵敏度分别为55.1%、69.6%和69.6%。在Ct<30的样本中，PrimeDirect、PrimeScript和Sansure检测方法的灵敏度分别提高到91.9%（n = 34/37）、89.2%（n = 33/37）和94.6%（n = 35/37）。PrimeDirect、PrimeScript和Sansure检测方法的特异性分别为88%、100%和100%。在本研究中，我们发现免提取PCR检测方法具有良好的灵敏度，尤其是对于高病毒载量（Ct<30）的样本，除了PrimeDirect检测方法的灵敏度和特异性不理想。尽管对于低病毒载量的样本灵敏度较低，但免提取试剂可为SARS-CoV-2诊断提供一种有价值的选择，更便宜、更简便且试剂消耗更少，尤其适用于分子检测经验较少和设备较差的实验室。
