Evaluation of the RealStar® SARS-CoV-2 RT-PCR kit RUO performances and limit of detection

Benoit Visseaux; Quentin Le Hingrat; Gilles Collin; Valentine Ferré; Alexandre Storto; Houria Ichou; Donia Bouzid; Nora Poey; Etienne de Montmollin; Diane Descamps; Nadhira Houhou-Fidouh

doi:10.1016/j.jcv.2020.104520

Evaluation of the RealStar® SARS-CoV-2 RT-PCR kit RUO performances and limit of detection

J Clin Virol. 2020 Aug:129:104520. doi: 10.1016/j.jcv.2020.104520. Epub 2020 Jun 29.

Affiliations

¹ Université de Paris, Assistance Publique - Hôpitaux de Paris, Service de virologie, Hôpital Bichat, Paris, France; UMR 1137-IAME, DeSCID: Decision SCiences in Infectious Diseases Control and Care, INSERM, Université de Paris, Paris, France. Electronic address: [email protected].
² Université de Paris, Assistance Publique - Hôpitaux de Paris, Service de virologie, Hôpital Bichat, Paris, France; UMR 1137-IAME, DeSCID: Decision SCiences in Infectious Diseases Control and Care, INSERM, Université de Paris, Paris, France.
³ Université de Paris, Assistance Publique - Hôpitaux de Paris, Service de virologie, Hôpital Bichat, Paris, France.
⁴ UMR 1137-IAME, DeSCID: Decision SCiences in Infectious Diseases Control and Care, INSERM, Université de Paris, Paris, France; Université de Paris, Assistance Publique - Hôpitaux de Paris, Service d'Accueil des Urgences, Hôpital Bichat, Paris, France.
⁵ Université de Paris, Assistance Publique - Hôpitaux de Paris, Service des Maladies Infectieuses et Tropicales, Hôpital Bichat, Paris, France.
⁶ Université de Paris, Assistance Publique - Hôpitaux de Paris, Réanimation Médicale et Infectieuses, Hôpital Bichat, Paris, France.

Abstract

Background: The use of efficient, reliable and sensitive PCR assays is a cornerstone in the race to contain the SARS-CoV-2 pandemic. In this work we performed an independent evaluation of the RealStar® SARS-CoV-2 RT-PCR Kit Researh Use Only (Altona) for SARS-CoV-2 detection.

Methods: A comparative limit of detection (LoD) assessment was performed between RealStar® SARS-CoV-2 and the currently WHO recommended RT-PCR (WHO-PCR) workflow using a quantified clinical sample. Assessment of the RealStar® SARS-CoV-2 assay was also performed using 83 primary clinical samples in comparison with the WHO-PCR.

Results: The RealStar® SARS-CoV-2 demonstrated a slightly higher sensitivity than the WHO recommended assay with a limit of detection at 625 copies/mL instead of 1250 copies/mL for the WHO-PCR in our conditions. The overall percent agreement between RealStar® SARS-CoV-2 and WHO-PCR on 83 clinical samples was 97.6 % (81/83) with a sensitivity at 97.8 % (45/46) and specificity at 97.3 % (36/37). No cross reaction was encountered for the other human coronaviruses (HKU1, OC43, NL63, 229E).

Conclusions: In this comparison of the RealStar® SARS-CoV-2 assay with the reference WHO assay, we observed a slightly better sensitivity of the RealStar® assay. It provides a robust option for all molecular biology laboratories, with a strong real-life LoD and is compatible with various real-time PCR platforms.

Keywords: COVID-19; RT-PCR; SARS-CoV-2; Sensitivity.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Betacoronavirus / isolation & purification*
COVID-19
COVID-19 Testing
COVID-19 Vaccines
Clinical Laboratory Techniques / methods*
Coronavirus Infections / diagnosis*
Humans
Limit of Detection
Molecular Diagnostic Techniques / methods*
Pandemics
Pneumonia, Viral / diagnosis*
Reverse Transcriptase Polymerase Chain Reaction / methods*
SARS-CoV-2
Sensitivity and Specificity