Colorimetric RT‑LAMP SARS‑CoV‑2 diagnostic sensitivity relies on color interpretation and viral load

dc.creatorAoki, Mateus Nóbrega
dc.creatorCoelho, Bruna de Oliveira
dc.creatorGóes, Luiz Gustavo Bentim
dc.creatorMinoprio, Paola
dc.creatorDurigon, Edison Luiz
dc.creatorMorello, Luis Gustavo
dc.creatorMarchini, Fabrício Klerynton
dc.creatorRiediger, Irina Natassja
dc.creatorDebur, Maria do Carmo
dc.creatorNakaya, Helder I.
dc.creatorBlanes, Lucas
dc.date2021-05-05T19:14:54Z
dc.date2021-05-05T19:14:54Z
dc.date2021
dc.date.accessioned2023-09-26T21:01:30Z
dc.date.available2023-09-26T21:01:30Z
dc.identifierAOKI, 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.
dc.identifier2045-2322
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/47062
dc.identifier10.1038/s41598-021-88506-y
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8867909
dc.descriptionThe 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.
dc.formatapplication/pdf
dc.languagepor
dc.publisherNature
dc.rightsopen access
dc.subjectCOVID-19
dc.subjectRT-LAMP
dc.subjectLoop Mediated Isothermal Amplifcation
dc.subjectReverse Transcriptase
dc.subjectCoronavirus Infections
dc.subjectReagent Kits, Diagnostic
dc.subjectPoint-of-Care Testing
dc.subjectADN Polimerasa Dirigida por ARN
dc.subjectInfecciones por Coronavirus
dc.subjectJuego de Reactivos para Diagnóstico
dc.subjectPruebas en el Punto de Atención
dc.subjectRNA-directed DNA polymerase
dc.subjectInfections à coronavirus
dc.subjectTrousses de réactifs pour diagnostic
dc.subjectAnalyse sur le lieu d'intervention
dc.subjectTranscriptase Reversa
dc.subjectInfecções por Coronavirus
dc.subjectKit de Reagentes para Diagnóstico
dc.subjectTestes Imediatos
dc.titleColorimetric RT‑LAMP SARS‑CoV‑2 diagnostic sensitivity relies on color interpretation and viral load
dc.typeArticle


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