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Comparison of In-house SARS-CoV-2 Genome Extraction Procedures. A Need for COVID-19 Pandemic

Overview
Journal J Virol Methods
Specialty Microbiology
Date 2021 Dec 13
PMID 34902458
Citations 1
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Abstract

Among the methods used to diagnose COVID-19, those based on genomic detection by q(RT)-PCR are the most sensitive. To perform these assays, a previous genome extraction of the sample is required. The dramatic increase in the number of SARS-CoV-2 detection assays has increased the demand for extraction reagents hindering the supply of commercial reagents. Homemade reagents and procedures could be an alternative. Nasopharyngeal samples were extracted by seven different methods as well as the automatic method MagNaPure96, to detect SARS-CoV-2. All protocols show sensitivity higher than 87 %, in comparison with reference method, for detecting SARS-CoV-2 as well as human β- globin. Our results support that these procedures, using common and cheap reagents, are effective to extract RNA (from SARS-CoV-2) or DNA (from human β-globin) genome from nasopharyngeal swabs. Furthermore, these procedures could be easily adopted by routine diagnostic laboratories to implement detection methods to help to fight against COVID-19 pandemic.

Citing Articles

Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use.

Bottino P, Zanotto E, Sidoti F, Pastrone L, Piva R, Mereu E Microorganisms. 2023; 11(2).

PMID: 36838500 PMC: 9960603. DOI: 10.3390/microorganisms11020535.

References
1.
Rodriguez A, Duyvejonck H, van Belleghem J, Gryp T, Van Simaey L, Vermeulen S . Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells. PLoS One. 2020; 15(2):e0229423. PMC: 7034890. DOI: 10.1371/journal.pone.0229423. View

2.
Verheyen J, Kaiser R, Bozic M, Timmen-Wego M, Maier B, Kessler H . Extraction of viral nucleic acids: comparison of five automated nucleic acid extraction platforms. J Clin Virol. 2012; 54(3):255-9. DOI: 10.1016/j.jcv.2012.03.008. View

3.
Wang C, Horby P, Hayden F, Gao G . A novel coronavirus outbreak of global health concern. Lancet. 2020; 395(10223):470-473. PMC: 7135038. DOI: 10.1016/S0140-6736(20)30185-9. View

4.
Arruda E, Hayden F . Detection of human rhinovirus RNA in nasal washings by PCR. Mol Cell Probes. 1993; 7(5):373-9. DOI: 10.1006/mcpr.1993.1055. View

5.
Yamada O, Matsumoto T, Nakashima M, Hagari S, KAMAHORA T, Ueyama H . A new method for extracting DNA or RNA for polymerase chain reaction. J Virol Methods. 1990; 27(2):203-9. DOI: 10.1016/0166-0934(90)90136-4. View