RT-LAMP Multicenter Study for SARS-CoV-2 Genome Molecular Detection in Brazilian Swab and Saliva Samples.

Autor: da Costa VD; Brazilian Reference Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., Santos AC; Brazilian Reference Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., da Silva LL; Brazilian Reference Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., Wiggers WJ; Service of Gastroenterology, Hepatology and Liver Transplantation, Hospital Nossa Senhora das Graças, Curitiba 80810-040, Brazil., Ivantes CAP; Service of Gastroenterology, Hepatology and Liver Transplantation, Hospital Nossa Senhora das Graças, Curitiba 80810-040, Brazil., Lima DM; Graduate Program in Medical Sciences, University of Fortaleza, Fortaleza 60811-905, Brazil., Colares JKB; Graduate Program in Medical Sciences, University of Fortaleza, Fortaleza 60811-905, Brazil., Dallacqua DSV; Molecular Virology Laboratory, Oswaldo Cruz Foundation, FIOCRUZ, Porto Velho 76812-245, Brazil., Motta-Castro ARC; Federal Faculty, University of Mato Grosso do Sul, Campo Grande 79070-900, Brazil., Dávila AMR; Computational and Systems Biology Laboratory, Graduate Program in Biodiversity and Health, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., Teles SA; Federal Faculty, University of Goiás, Goiânia 74690-900, Brazil., Carneiro MADS; Federal Faculty, University of Goiás, Goiânia 74690-900, Brazil., Caetano KAA; Federal Faculty, University of Goiás, Goiânia 74690-900, Brazil., Anunciação FAC; Hospital Getúlio Vargas, Teresina 64001-020, Brazil.; Natan Portela Institute of Tropical Diseases, Teresina 64002-510, Brazil., de Paula VS; Molecular Virology Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., Villar LM; Brazilian Reference Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-360, Brazil., On Behalf Of The Brazilian Covid-Research Group
Jazyk: angličtina
Zdroj: Diagnostics (Basel, Switzerland) [Diagnostics (Basel)] 2023 Jan 06; Vol. 13 (2). Date of Electronic Publication: 2023 Jan 06.
DOI: 10.3390/diagnostics13020210
Abstrakt: Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a rapid method that can replace RT-qPCR. A simple molecular assay for SARS-CoV-2 RNA detection in gold-standard diagnosis through swabs and alternative specimens such as saliva could be helpful in promoting genomic surveillance. A multicenter study was conducted to evaluate the RT-LAMP assay method as an alternative for the molecular detection of SARS-CoV-2 lineages in swab and saliva samples. A total of 350 swabs from individuals with ( n = 276) or without ( n = 74) COVID-19 tested by RT-qPCR were collected. Paired saliva was also collected from 90 individuals who had SARS-CoV-2 RNA that was detectable ( n = 30) or undetectable ( n = 60) via RT-qPCR. For the RT-LAMP methodology, six primers were used for ORF1 gene amplification. As for SARS-CoV-2 genotyping, 39 swabs had the whole genome sequenced by MinION. The sensitivity of RT-LAMP to the swab was 90.2%. For the swab samples with Ct ≤ 30, the sensitivity improved by 96%. Considering saliva with Ct ≤ 30 in RT-qPCR testing, the RT-LAMP sensitivity was 100%. The RT-LAMP specificity was 100% for both the swab and saliva samples. This RT-LAMP assay was capable of detecting all the SARS-CoV-2 lineages circulating in the Brazilian swab samples. The RT-LAMP method has significant potential for use in clinical routines since it was capable of detecting SARS-CoV-2 RNA in swab and saliva samples.
Databáze: MEDLINE
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