NEOKIT-COVID19, a colorimetric reverse-transcription loop-mediated isothermal amplification test to detect sars-cov-2 and its variants from extraction-free samples

WERBAJH S; LAROCCA, L; CARRILLO C; STOLOWICS FG; OGAS, L; PALLOTO, S; CASSARA, S; ZAPIOLA, I; BOAUZAS MB; VOJNOV A

Congreso; REUNIÓN DE SOCIEDADES DE BIOCIENCIAS 2021; 2021

We present a colorimetric SARS-CoV-2 RNA detection method performed using RT-LAMP to achieve specific, rapid, with a detection limit of 25-100 copies per reaction directly from NP or saliva samples, without RNA isolation. We optimized the final condition of temperature (64°C), time (60m) and sample pretreatment with LB with a heat inactivation of 8 minutes. The assay present 90,6 % of sensitivity and 100 % of specificity.The kit, NEOKIT-PLUS, was authorized by ANMAT, Argentina, after validating it using samples previously analyzed by RT-qPCR. The primer binding sites are well conserved in all the variants of concern (VOC), notified by World Health Organization (WHO). These lineages include B.1.1.7, B.1.351, P.1 and B.1.617.2.