A two minute liquid based sample preparation for rapid SARS-CoV2 real-time PCR screening: A multicentre evaluation

Eric C J Claas; Pieter W Smit; Mario J A W M van Bussel; Harold Verbakel; Mohammed Taouil; Jaco J Verweij; Steven F T Thijsen

doi:10.1016/j.jcv.2020.104720

A two minute liquid based sample preparation for rapid SARS-CoV2 real-time PCR screening: A multicentre evaluation

J Clin Virol. 2021 Feb:135:104720. doi: 10.1016/j.jcv.2020.104720. Epub 2020 Dec 31.

Authors

Eric C J Claas¹, Pieter W Smit², Mario J A W M van Bussel³, Harold Verbakel⁴, Mohammed Taouil², Jaco J Verweij⁴, Steven F T Thijsen⁵

Affiliations

¹ Leiden University Medical Center, Department of Medical Microbiology, Leiden, the Netherlands. Electronic address: e.claas@lumc.nl.
² Maasstad Hospital, Medical Microbiology Laboratory, Rotterdam, the Netherlands.
³ Leiden University Medical Center, Department of Medical Microbiology, Leiden, the Netherlands.
⁴ Elisabeth-Tweesteden Hospital, Microvida Laboratory for Medical Microbiology and Immunology, Tilburg, the Netherlands.
⁵ Diakonessenhuis Hospital, Department of Medical Microbiology and Immunology, the Netherlands.

Abstract

Background: Apart from major health concerns associated to the SARS-coronavirus-2 (SARS-CoV-2) pandemic, also the diagnostic workflow encountered serious problems. Limited availability of kit components, buffers and even plastics has resulted in suboptimal testing procedures worldwide. Alternative workflows have been implemented to overcome these difficulties. Recently a liquid based sample prep has been launched as solution to overcome limitations in relation to nucleic acid extraction.

Objective: Multicenter evaluation of the QIAprep&amp Viral RNA UM kit (QIA P&A) for rapid sample preparation and real-time PCR detection of SARS-CoV-2 in comparison to standardized laboratory testing methods.

Study design: Selected samples of the routine diagnostic workflow at Clinical Microbiology Laboratories of four Dutch hospitals have been subjected to the rapid QIA P&A protocol and the results have been compared to routine diagnostic data.

Results: Combining results of manual and automated procedures, a total of 377 clinical samples of which 202 had been tested positive with a wide range of C_T values, showed almost complete concordance in the QIA P&A assay for samples up to C_T values of 33 with one exception of C_T 31. Prospectively 60 samples were tested and also showed 100 % concordance with 5 positives. The method has been automated by two centres.

Conclusions: Despite an input of only 8 μL of clinical sample, the QIA P&A kit showed good performance for sample preparation and amplification of SARS-CoV-2 and can contribute as a rapid molecular testing strategy in managing the CoV-2 pandemic.

Keywords: Real-time PCR; SARS-CoV-2; Sample preparation.

Publication types

Multicenter Study

MeSH terms

COVID-19 / diagnosis*
COVID-19 / virology*
COVID-19 Testing / methods*
Clinical Laboratory Techniques / methods
Humans
Mass Screening / methods*
Molecular Diagnostic Techniques / methods
Pandemics / prevention & control
Prospective Studies
Real-Time Polymerase Chain Reaction / methods*
SARS-CoV-2 / genetics*
Specimen Handling / methods
Workflow