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Preprint / Version 1

Real-time PCR-based SARS-CoV-2 detection

##article.authors##

José Francisco Cancino Mesa Granma Medical Science University, Manzanillo Medical Faculty Celia Sánchez Manduley, Cuba. https://orcid.org/0000-0002-9328-9856
Adrián Alejandro Vitón Castillo Pinar del Río Medical Science University, Ernesto Guevara de la Serna Medical Faculty, Pinar del Río, Cuba https://orcid.org/0000-0002-7811-2470

DOI:

https://doi.org/10.1590/SciELOPreprints.707

Keywords:

Polymerase Chain Reaction, Coronavirus Infections, Clinical Laboratory Techniques

Abstract

Introduction: reverse transcription-polymerase chain reaction (RT-PCR) is an accurate technique in coronavirus detection. It was selected as a confirmatory test for SARS-CoV-2 infection due to proven safety in the diagnosis of MERS-CoV and SARS-CoV strains.

Objective: to characterize the reverse-transcription polymerase chain reaction assay as the main diagnostic test used to confirm SARS-CoV-2 infection during the active period of viral replication.

Method: a literature review was conducted in articles published up to May 2020. The following databases were consulted: Scopus, Wiley Online Library, SciELO, DIALNET, EBSCO, MEDLINE, and PubMed. Articles in Spanish and English were retrieved, selecting 46 references.

Development: reverse transcription-polymerase chain reaction (RT-PCR) assay to detect SARS-CoV-2, targets to ORF1ab fragments, RNA polymerase dependent (RdRp), the E gene, the N gene, and the S gene. Nasopharyngeal swab proved to achieve better results than oropharyngeal swab and saliva. RT-PCR testing using rectal swab specimens appeared required on suspected false-negative cases. The Reverse Transcription Loop-mediated Isothermal Assay showed faster diagnosis compared to RT-PCR with equal sensitivity and specificity.

Conclusions: the availability of diagnostic tests is crucial for the isolation of positive cases and tracking the transmission epidemiological chain. RT-PCR proved to be the test of choice during the active period of viral replication. The RT-LAMP assay is a rapid diagnostic alternative with similar principles to the RT-PCR technique.