Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection.

Autor: Barza R; NorthShore University HealthSystem, Department of Pathology and Laboratory Medicine, Evanston, IL, USA., Patel P; NorthShore University HealthSystem, Department of Pathology and Laboratory Medicine, Evanston, IL, USA., Sabatini L; NorthShore University HealthSystem, Department of Pathology and Laboratory Medicine, Evanston, IL, USA., Singh K; NorthShore University HealthSystem, Department of Pathology and Laboratory Medicine, Evanston, IL, USA. Electronic address: ksingh@northshore.org.
Jazyk: angličtina
Zdroj: Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology [J Clin Virol] 2020 Nov; Vol. 132, pp. 104587. Date of Electronic Publication: 2020 Aug 11.
DOI: 10.1016/j.jcv.2020.104587
Abstrakt: The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. RT-PCR results using the ChromaCode HDPCR™ SARS-CoV-2 were compared using the heat-RNA release method and an automated RNA extraction system (EMAG). The heat-RNA release method correctly identified 94 % (81/86 nasopharyngeal samples) that were positive for SARS-CoV-2. Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: