Estandarización y validación de una prueba molecular RT-LAMP in house para el diagnóstico de SARS-CoV-2

Oscar Escalante-Maldonado, Margot Vidal-Anzardo, Fernando Donaires, Gilmer Solis-Sanchez, Italo Gallesi, Luis Pampa-Espinoza, Maribel Huaringa, Nancy Rojas-Serrano, Coralith García, Eddie Angles-Yanqui, Ronnie Gustavo Gavilán, Ricardo Durães-Carvalho, Jairo Mendez-Rico, César Cabezas, Paulo Vitor Marques-Simas

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

Resumen

OBJECTIVES: To standardize and validate an in-house RT-LAMP test for the detection of SARS-CoV-2, based on laboratory and field assays using samples from COVID-19 suspected patients. MATERIALS AND METHODS: An in-house SARS-CoV-2 RT-LAMP molecular test was standardized, establishing the detection limit with Vero cells of isolated Peruvian strains of SARS-CoV-2, and the robustness to various concentrations of primers. The laboratory validation was performed with 384 nasal and pharyngeal swab samples (UFH) obtained between March and July 2020. The field validation was performed with 383 UFH obtained from COVID-19 suspected symptomatic cases. All samples were tested by RT-LAMP and RT-qPCR. The RT-qPCR was considered as the reference standard test. The concordance measures and diagnostic performance were calculated. RESULTS: The detection limit was consistent in cases with Ct <30 in both tests, showing efficiency to detect up to 1000 copies/μL of the target gene. Robustness was evidenced with half of the primer concentrations and 20 μL of final volume. Absence of amplification was identified for other HCoVs. Concordance showed a kappa index of 0.88 (95% CI: 0.83-0.93) and 0.89 (95% CI: 0.84 - 0.94) in laboratory and field settings, respectively. The sensitivity value in the laboratory was 87.4% (95% CI: 80.8 - 92.4) and 88.1% in the field (95% CI: 81.6 - 92.9). The specificity value in both settings was 98.8% (95% CI: 96.4-99.7). CONCLUSIONS: The in-house SARS-CoV-2 RT-LAMP test was successfully validated based on its adequate robustness, no cross-reactions, good concordance, and diagnostic performance compared to RT-qPCR.

Título traducido de la contribuciónStandardization and validation of an in house RT-LAMP molecular test for the diagnosis of SARS-CoV-2
Idioma originalEspañol
Páginas (desde-hasta)7-16
Número de páginas10
PublicaciónRevista Peruana de Medicina de Experimental y Salud Publica
Volumen38
N.º1
DOI
EstadoPublicada - 1 ene 2021

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