Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 291, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2021.114086
Keywords
SARS-CoV-2; RT-PCR; Extraction-Free
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The study evaluated the performance of different extraction-free SARS-CoV-2 RT-PCR assays, showing good sensitivity, especially for high viral loads (Ct<30), with PrimeScript and Sansure assays performing well.
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 RTPCR. 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.
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