Article
Colorimetric RT‑LAMP SARS‑CoV‑2 diagnostic sensitivity relies on color interpretation and viral load
Registro en:
AOKI, Mateus Nóbrega et al. Colorimetric RT‑LAMP SARS‑CoV‑2 diagnostic sensitivity relies on color interpretation and viral load. Scientific Reports, v.11, n. 9026, p. 1–10, 2021.
2045-2322
10.1038/s41598-021-88506-y
Autor
Aoki, Mateus Nóbrega
Coelho, Bruna de Oliveira
Góes, Luiz Gustavo Bentim
Minoprio, Paola
Durigon, Edison Luiz
Morello, Luis Gustavo
Marchini, Fabrício Klerynton
Riediger, Irina Natassja
Debur, Maria do Carmo
Nakaya, Helder I.
Blanes, Lucas
Resumen
The use of RT-LAMP (reverse transcriptase—loop mediated isothermal amplifcation) has been considered as a promising point-of-care method to diagnose COVID-19. In this manuscript we show that the RT-LAMP reaction has a sensitivity of only 200 RNA virus copies, with a color change from pink to yellow occurring in 100% of the 62 clinical samples tested positive by RT-qPCR. We also demonstrated that this reaction is 100% specifc for SARS-CoV-2 after testing 57 clinical samples infected with dozens of diferent respiratory viruses and 74 individuals without any viral infection. Although the majority of manuscripts recently published using this technique describe only the presence of two-color states (pink =negative and yellow=positive), we verifed by naked-eye and absorbance measurements that there is an evident third color cluster (orange), in general related to positive samples with low viral loads, but which cannot be defned as positive or negative by the naked eye. Orange colors should be repeated or tested by RT-qPCR to avoid a false diagnostic. RT-LAMP is
therefore very reliable for samples with a RT-qPCR Ct < 30 being as sensitive and specifc as a RT-qPCR
test. All reactions were performed in 30 min at 65 °C. The use of reaction time longer than 30 min is also not recommended since nonspecifc amplifcations may cause false positives.