Detección del virus SARS-CoV-2 a través de la prueba de amplificación isotérmica mediada por bucle con transcripción reversa (RT-LAMP)
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https://doi.org/10.54104/saywa.v5n6.1506Palabras clave:
COVID-19, RT-LAMP, SARS-CoV-2Resumen
Resumen
El propósito del presente estudio fue estandarizar una metodología rápida y de simple ejecución para la detección de SARS-CoV-2, usando la técnica de amplificación de ácidos nucleicos mediada por bucle con transcripción inversa en condiciones isotérmicas (RT-LAMP, del inglés Reverse Transcription Loop-mediated isothermal amplification), para el diagnóstico de la COVID-19. La estandarización se realizó en muestras de ARN previamente confirmadas por RT-qPCR usando el protocolo Charité, Berlín, Alemania. Detallamos los pasos de la técnica y destacamos el uso del cebador S-123. En conclusión, la prueba RT-LAMP con el set de cebadores S-123 para SARS-CoV-2 en muestras de ARN, mostró un alto desempeño con relación a la técnica gold standard RT-qPCR.
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P. L. Hurtado L, Díaz D, Escorcia K, Flórez L, Bello Y, Díaz Y, Navarro E, Pacheco LC, Galán N, Maestre R, Acosta A, “Validación clínica de la prueba RT-LAMP para el diagnóstico rápido del SARS-CoV-2. biomedica,” Biomed. [Internet], vol. 42, 2022. https://revistabiomedica.org/index.php/biomedica/article/view/6523
A. A. Artik Y., Coşğun A.B., Cesur N.P., Hızel N., Uyar Y., Sur H., “Comparison of COVID-19 laboratory diagnosis by commercial kits: Effectivity of RT-PCR to the RT-LAMP,” J. Med. Virol., pp. 1–10, 2022. https://doi.org/10.1002/jmv.27559
F. A. Méndez Tibambre, M. E., Rodríguez Parra, Z.J., Portela Dussan, D. D., Bustamante Restrepo, M.F., Aristizábal Gutiérrez, “Método LAMP como alternativa diagnóstica para la detección del virus SARS-CoV-2,” Rev. Colomb. Ciencias Químico-Farmacéuticas, vol. 50, no. 3, 2021. https://doi.org/10.15446/rcciquifa.v50n3.100204
J. El-Tholoth, M., Bau, H. H., & Song, “A Single and Two-Stage, Closed-Tube, Molecular Test for the 2019 Novel Coronavirus (COVID-19) at Home, Clinic, and Points of Entry,” ChemRxiv Prepr. Serv. Chem., 2020. https://doi.org/10.26434/chemrxiv.11860137
B. L. Lamers, M.M., Haagmans, “SARS-CoV-2 pathogenesis,” Nat Rev Microbiol, vol. 20, pp. 270–284, 2022. https://doi.org/10.1038/s41579-022-00713-0
T. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., & Hase, “Loop-mediated isothermal amplification of DNA,” Nucleic Acids Res., vol. 28, no. 12, p. E63, 2000. https://doi.org/10.1093/nar/28.12.e63
M. J. Heithoff, D. M., Barnes, L., 5th, Mahan, S. P., Fox, G. N., Arn, K. E., Ettinger, S. J., Bishop, A. M., Fitzgibbons, L. N., Fried, J. C., Low, D. A., Samuel, C. E., & Mahan, “Assessment of a Smartphone-Based Loop-Mediated Isothermal Amplification Assay for Detection of SARS-CoV-2 and Influenza Viruses,” JAMA Netw. open, vol. 5, no. 1, 2022. https://doi.org/10.1001/jamanetworkopen.2021.45669
B. A. Londono-Avendano, M.A., Libreros, G., Osorio, L., Parra, “Rapid RT-LAMP Assay for SARS-CoV-2 with Colorimetric Detection Assisted by a Mobile Application,” Diagnostics, vol. 12, 2022. https://doi.org/10.3390/diagnostics12040848
X. et al Boni, M.F., Lemey, P., Jiang, “Evolutionary origins of the SARS-CoV-2 sarbecovirus lineage responsible for the COVID-19 pandemic,” Nat. Microbiol., vol. 5, pp. 1408–1417, 2020. https://doi.org/10.1038/s41564-020-0771-4
J. Yan, C., Cui, J., Huang, L., Du, B., Chen, L., Xue, G., Li, S., Zhang, W., Zhao, L., Sun, Y., Yao, H., Li, N., Zhao, H., Feng, Y., Liu, S., Zhang, Q., Liu, D., & Yuan, “Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay,” Clin. Microbiol. Infect. Off. Publ. Eur. Soc. Clin. Microbiol. Infect. Dis., vol. 26, no. 6, pp. 773–779, 2020. https://doi.org/10.1016/j.cmi.2020.04.001
X. Huang, X., Tang, G., Ismail, N., Wang, “Developing RT-LAMP assays for rapid diagnosis of SARS-CoV-2 in saliva,” eBioMedicine, vol. 75, pp. 1–15, 2022. https://doi.org/10.1016/j.ebiom.2021.103736
Y. A. Kashir. J, “Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19,” Med. Hypotheses, vol. 141, 2020. https://doi.org/10.1016/j.mehy.2020.109786.
M. M. M. Moreno-Soto A.A., Espinoza Venegas L.A., Siles Montoya C.A, “Coinfección de la COVID 19 y Virus del Dengue:Reporte de Caso,” Acta Médica Peru., vol. 38, no. 2, pp. 123–126, 2021. https://doi.org/10.35663/amp.2021.382.2031
Y. Z. Wu,F., Zhao,S., Yu,B., Chen,Y.M., Wang,W., Song,Z.G., Hu,Y., Tao,Z.W., Tian,J.H., Pei,Y.Y., Yuan,M.L., Zhang,Y.L., Dai,F.H., Liu,Y., Wang,Q.M., Zheng,J.J., Xu,L., Holmes,E.C. and Zhang, “A new coronavirus associated with human respiratory disease in China,” Nature, vol. 579, no. 7798, pp. 265–269, 2020. https://www.nature.com/articles/s41586-020-2008-3
Z. Huang, W. E., Lim, B., Hsu, C. C., Xiong, D., Wu, W., Yu, Y., Jia, H., Wang, Y., Zeng, Y., Ji, M., Chang, H., Zhang, X., Wang, H., & Cui, “RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2,” Microb. Biotechnol., vol. 13, no. 4, pp. 950–961, 2020. https://doi.org/10.1111/1751-7915.13586
E. et al Amaral, C., Antunes, W., Moe, “A molecular test based on RT-LAMP for rapid, sensitive and inexpensive colorimetric detection of SARS-CoV-2 in clinical samples,” Sci Rep, vol. 11, no. 16430, 2021. https://doi.org/10.1038/s41598-021-95799-6
P. L. Silva SJRD, Pardee K, “Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review,” Viruses, vol. 12, no. 1, 2019. https://pubmed.ncbi.nlm.nih.gov/31877989/
T. N. Zhang Y, “Improving RT-LAMP detection of SARS-CoV-2 RNA through primer set selection and combination,” PLoS One, vol. 17, no. 4, 2022. https://doi.org/10.1371/journal.pone.0254324
L. G. B. et al. Aoki, M.N., de Oliveira Coelho, B., Góes, “Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load,” Sci Rep, vol. 11, no. 9026, 2021. https://doi.org/10.1038/s41598-021-88506-y
D. A. et al. Lim, B., Ratcliff, J., Nawrot, “Clinical validation of optimised RT-LAMP for the diagnosis of SARS-CoV-2 infection,” Sci Rep, vol. 11, no. 16193, 2021. https://doi.org/10.1038/s41598-021-95607-1
A. Reynés, B., Serra, F., Palou, “Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification,” Biotechniques, vol. 70, pp. 219–225, 2021. https://doi.org/10.2144/btn-2020-0159
J. Lalli, M. A., Langmade, S. J., Chen, X., Fronick, C. C., Sawyer, C. S., Burcea, L. C., Wilkinson, M. N., Fulton, R. S., Heinz, M., Buchser, W. J., Head, R. D., Mitra, R. D., & Milbrandt, “Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP,” medRxiv, 2021. https://doi.org/10.1101/2020.05.07.20093542
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Derechos de autor 2024 Elizabeth Ruiz Marvez, Ana Luisa Muñoz Ramírez, Orlando Alfredo Torres García, Francisco Javier Vargas Ortíz, Juana Patricia Sánchez Villamil
Esta obra está bajo una licencia internacional Creative Commons Atribución-NoComercial-CompartirIgual 4.0.
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