Publicación:
A Multiplex and Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Sensitive and Rapid Detection of Novel SARS-CoV-2
A Multiplex and Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Sensitive and Rapid Detection of Novel SARS-CoV-2
| dc.contributor.author | Juscamayta-López, E. | es_PE |
| dc.contributor.author | Valdivia, F. | es_PE |
| dc.contributor.author | Horna, H. | es_PE |
| dc.contributor.author | Tarazona, D. | es_PE |
| dc.contributor.author | Linares, L. | es_PE |
| dc.contributor.author | Rojas, N. | es_PE |
| dc.contributor.author | Huaringa, M. | es_PE |
| dc.date.accessioned | 2024-05-30T23:13:38Z | |
| dc.date.available | 2024-05-30T23:13:38Z | |
| dc.date.issued | 2021 | |
| dc.description | This work was supported by the CONCYTEC-FONDECYT Program of Proyectos Especiales: Respuesta al COVID-19 2020-01-01 [grant number 034-2020-FONDECYT] and the National Institute of Health of Peru. | |
| dc.description.abstract | Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a major threat to public health. Rapid molecular testing for convenient and timely diagnosis of SARS-CoV-2 infections represents a challenge that could help to control the current pandemic and prevent future outbreaks. We aimed to develop and validate a multiplex and colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay using lyophilized LAMP reagents for sensitive and rapid detection of SARS-CoV-2. LAMP primers were designed for a set of gene targets identified by a genome-wide comparison of viruses. Primer sets that showed optimal features were combined into a multiplex RT-LAMP assay. Analytical validation included assessment of the limit of detection (LoD), intra- and inter-assay precision, and cross-reaction with other respiratory pathogens. Clinical performance compared to that of real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) was assessed using 278 clinical RNA samples isolated from swabs collected from individuals tested for COVID-19. The RT-LAMP assay targeting the RNA-dependent RNA polymerase (RdRp), membrane (M), and ORF1ab genes achieved a comparable LoD (0.65 PFU/mL, CT=34.12) to RT-qPCR and was 10-fold more sensitive than RT-qPCR at detecting viral RNA in clinical samples. Cross-reactivity to other respiratory pathogens was not observed. The multiplex RT-LAMP assay demonstrated a strong robustness and acceptable intra- and inter-assay precision (mean coefficient of variation, 4.75% and 8.30%). Diagnostic sensitivity and specificity values were 100.0% (95% CI: 97.4–100.0%) and 98.6% (95% CI: 94.9–99.8%), respectively, showing high consistency (Cohen’s kappa, 0.986; 95% CI: 0.966–1.000; p<0.0001) compared to RT-qPCR. The novel one-step multiplex RT-LAMP assay is storable at room temperature and showed similar diagnostic accuracy to conventional RT-qPCR, while being faster (<45 min), simpler, and cheaper. The new assay could allow reliable and early diagnosis of SARS-CoV-2 infections in primary health care. It may aid large-scale testing in resource-limited settings, especially if it is integrated into a point-of-care diagnostic device. © Copyright © 2021 Juscamayta-López, Valdivia, Horna, Tarazona, Linares, Rojas and Huaringa. | |
| dc.description.sponsorship | Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - Concytec | |
| dc.identifier.doi | https://doi.org/10.3389/fcimb.2021.653616 | |
| dc.identifier.scopus | 2-s2.0-85110180024 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12390/2960 | |
| dc.language.iso | eng | |
| dc.publisher | Frontiers Media S.A. | |
| dc.relation.ispartof | Frontiers in Cellular and Infection Microbiology | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject | timely diagnosis | |
| dc.subject | COVID-19 | es_PE |
| dc.subject | diagnostic accuracy | es_PE |
| dc.subject | Multiplex RT-LAMP | es_PE |
| dc.subject | primary health care | es_PE |
| dc.subject | rapid molecular tests | es_PE |
| dc.subject | resource-limited settings | es_PE |
| dc.subject | SARS-CoV-2 | es_PE |
| dc.subject.ocde | https://purl.org/pe-repo/ocde/ford#3.03.08 | |
| dc.title | A Multiplex and Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Sensitive and Rapid Detection of Novel SARS-CoV-2 | |
| dc.type | info:eu-repo/semantics/article | |
| dspace.entity.type | Publication |