Alternative RNA extraction-free techniques for the real-time RT-PCR detection of SARS-CoV-2 in nasopharyngeal swab and sputum samples

Stephany D. Villota, Victoria E. Nipaz, Andrés Carrazco-Montalvo, Sarah Hernandez, Jesse J. Waggoner, Patricio Ponce, Josefina Coloma, Alberto Orlando, Varsovia Cevallos

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

6 Citas (Scopus)

Resumen

Standard diagnoses of SARS-CoV-2 infections are done by RNA extraction and real-time RT-PCR (rRT-PCR). However, the need for RNA extraction complicates testing due to increased processing time, high cost, and limited availability of commercial kits. Therefore, alternative methods for rRT-PCR detection of SARS-CoV-2 without RNA extraction were investigated. Nasopharyngeal and sputum samples were used to compare the sensitivity of three techniques: Trizol RNA extraction, thermal shock, and the direct use of samples with an RNase inhibitor. Direct, extraction-free use of primary samples plus the RNase inhibitor produced diagnostic values of 100 % sensitivity and specificity compared to standard protocols, and these findings were validated in a second, independent laboratory.

Idioma originalInglés
Número de artículo114302
PublicaciónJournal of Virological Methods
Volumen298
DOI
EstadoPublicada - dic. 2021
Publicado de forma externa

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