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Método LAMP como alternativa diagnostica para la detección del virus SARS-CoV-2
LAMP method as a diagnostic alternative for the detection of the SARS-CoV-2 virus
Método LAMP como alternativa de diagnóstico para a detecção do vírus SARS-CoV-2
DOI:
https://doi.org/10.15446/rcciquifa.v50n3.100204Palabras clave:
Diagnóstico molecular, RT-LAMP, COVID-19, SARS-CoV-2 (es)Diagnóstico molecular, RT-LAMP, COVID-19, SARS-CoV-2 (pt)
Molecular diagnostic, RT-LAMP, COVID-19, SARS-CoV-2 (en)
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Introducción: el coronavirus SARS-CoV-2, causante de la enfermedad COVID-19 se ha convertido en un problema de salud pública mundial que requiere la implemen-tación de pruebas de diagnóstico rápidas y sensibles. Objetivo: evaluar y comparar el límite de detección del método LAMP con respecto al método estándar y aplicar el método RT-LAMP para el diagnóstico de SARS-CoV-2 en muestras clínicas de pacientes colombianos. Métodos: se realizó un estudio descriptivo y transversal, analizando un total de 25 muestras de hisopado nasofaríngeo incluyendo muestras negativas y positivas para SARS-CoV-2, a través del método RT-LAMP comparado con el método estándar RT-qPCR. Resultados: el método LAMP detectó ~18 copias del gen N, en 30 min, demostrando un límite de detección similar al obtenido con el método estándar, en un menor tiempo y una concordancia en RT-LAMP del 100 % con los resultados. Conclusiones: RT-LAMP es un método sensible, específico y rápido que puede ser empleado para el diagnóstico de la enfermedad COVID-19.
Introduction: The SARS-CoV-2 coronavirus, that causes the COVID-19 disease, has become a global public health problem that requires the implementation of rapid and sensitive diagnostic tests. Aim: To evaluate and compare the sensitivity of LAMP assay to a standard method and use RT-LAMP for the diagnosis of SARS-CoV-2 in clinical samples from Colombian patients. Methods: A descriptive and cross-sectional study was conducted. A total of 25 nasopharyngeal swab samples including negative and positive samples for SARS-CoV-2 were analyzed, through the RT-LAMP method compared to the RT-qPCR assay. Results: LAMP method detected ~18 copies of the N gene, in 30 min, evidenced a detection limit similar to the standard method, in a shorter time and a concordance in RT-LAMP of 100% with the results. Conclusions: RT-LAMP is a sensitive, specific, and rapid method that can be used as a diagnostic aid of COVID-19 disease.
Introdução: o coronavírus SARS-CoV-2, causador da doença de COVID-19, tornou-se um problema de saúde pública global que requer a implementação de testes diagnósticos rápidos e sensíveis. Objetivo: avaliar e comparar o limite de detecção do método LAMP em relação ao método padrão e aplicar o método RT-LAMP para o diagnóstico de SARS-CoV-2 em amostras clínicas de pacientes colombianos. Métodos: foi realizado um estudo descritivo e transversal, analisando um total de 25 amostras de hissopado nasofaríngeo, incluindo amostras negativas e positivas para SARS-CoV-2, pelo método RT-LAMP em comparação ao método RT-qPCR padrão. Resultados: o método LAMP detectou ~18 cópias do gene N, em 30 min., demonstrando um limite de detecção semelhante ao obtido com o método padrão, em um tempo menor e uma concordância no RT-LAMP de 100% com os resul-tados. Conclusões: RT-LAMP é um método sensível, específico e rápido que pode ser usado para o diagnóstico da doença de COVID-19.
Referencias
M.M. Oliva-Martínez, La COVID-19; pandemia del siglo XXI, MediMay,Revista de Ciencias Médicas de Mayabeque, 27(2), 99-103 (2020).
Instituto Nacional de Salud Ministerio de Salud y Protección social, Coronavi-rus (COVID-19Covid-19), URL: http://www.minsalud.gov.co/salud/publica/PET/Paginas/Covid-19_copia.aspx, consultado en noviembre de 2020.
Organización Mundial de la Salud (OMS), Coronavirus Disease (COVID-19) Dashboard, URL: https://covid19.who.int, consultado en noviembre de 2020.
B.S. Kamps, C. Hoffmann, Epidemiología. en: Covid Reference, Steinhaüser Ver-lag, España, 2020, pp. 1-111.
N. Zhu, D. Zhang, W. Wang, X. Li, B. Yang, J. Song., et al., A novel coronavi-rus from patients with pneumonia in China, New England Journal of Medicine, 382(8), 727-733 (2020). DOI: https://doi.org/10.1056/NEJMoa2001017
F. Wu, S. Zhao, B. Yu, Y.M. Chen, W. Wang, Z.G. Song, et al., A new coronavirus associated with human respiratory disease in China, Nature, 579(7798), 265-269 (2020). DOI: https://doi.org/10.1038/s41586-020-2008-3
M.L. Holshue, C. DeBolt, S. Lindquist, K.H. Lofy, J. Wiesman, H. Bruce, et al., First case of 2019 novel coronavirus in the United States, New England Journal of Medicine, 382, 929-936 (2020). DOI: https://doi.org/10.1056/NEJMoa2001191
B. Udugama, P. Kadhiresan, H.N. Kozlowski, A. Malekjahani, M. Osborne, V.Y.C. Li, et al., Diagnosing COVID-19: The disease and tools for detection, ACS nano, 14, 3822-3835 (2020). DOI: https://doi.org/10.1021/acsnano.0c02624
J.P. Broughton, X. Deng, G. Yu, C.L. Fasching, V. Servellita, J. Singh, et al., CRISPR–Cas12-based detection of SARS-CoV-2, Nature Biotechnolog y, 38, 870-874 (2020). DOI: https://doi.org/10.1038/s41587-020-0513-4
J. Kashir, A. Yaqinuddin, Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19, Medical Hypotheses, 141, 109786 (2020). DOI: https://doi.org/10.1016/j.mehy.2020.109786
V. Corman, M. Müller, U. Costabel, J. Timm, T. Binger, B. Meyer, et al., Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infec-tions, Euro Surveillance, 17(49), 20334 (2020). DOI: https://doi.org/10.2807/ese.17.49.20334-en
V. Corman, O. Landt, M. Kaiser, R. Molenkamp, A. Meijer, D.K.W. Chu, et al., Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR, Euro Surveillance, 25(3), 2000045 (2020). DOI: https://doi.org/10.2807/1560-7917.ES.2020.25.3.2000045
Universidad de la República Uruguay, Nuevo test diagnóstico de COVID-19, URL: http:// https://udelar.edu.uy/portal/2020/08, consultado en noviembre de 2020.
T. Notomi, H. Okayama, H. Masubuchi, T. Yonekawa, K. Watanabe, N. Amino, et al., Loop-mediated isothermal amplification of DNA, Nucleic Acids Research, 28(12), 63 (2000). DOI: https://doi.org/10.1093/nar/28.12.e63
J. Hill, S. Beriwal, I. Chandra, V.K. Paul, A. Kapil, T. Singh, et al., Loop-mediated isothermal amplification assay for rapid detection of common strains of Escheri-chia coli, Journal of Clinical Microbiolog y, 46(8), 2800-2804 (2008). DOI: https://doi.org/10.1128/JCM.00152-08
N. Tomita, Y. Mori, H. Kanda, T. Notomi, Loop-mediated isothermal amplifica-tion (LAMP) of gene sequences and simple visual detection of products, Nature Protocols, 3(5), 877-882 (2008). DOI: https://doi.org/10.1038/nprot.2008.57
W.E. Huang, B. Lim, C.C. Hsu, D. Xiong, W. Wu, Y. Yu, et al., RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2, Microbial Biotechnolog y, 13(4), 950-961 (2020). DOI: https://doi.org/10.1111/1751-7915.13586
M. Parida, S. Sannarangaiah, P. Dash, P.V.L. Rao, K. Morita, Loop mediated isothermal amplification (LAMP): A new generation of innovative gene amplifi-cation technique; perspectives in clinical diagnosis of infectious diseases, Reviews in Medical Virolog y, 18(6), 407-421 (2008). DOI: https://doi.org/10.1002/rmv.593
M. Goto, E. Honda, A. Ogura, A. Nomoto, K.I. Hanaki, Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue, Biotechniques, 46(3), 167-172 (2009). DOI: https://doi.org/10.2144/000113072
A. Srisrattakarn, A. Lulitanond, C. Wilailuckana, N. Charoensri, L. Won-glakorn, P. Saenjamla, et al., Rapid and simple identification of carbapenemase genes, bla NDM, bla OXA-48, bla VIM, bla IMP-14 and bla KPC groups, in Gram-negative bacilli by in-house loop-mediated isothermal amplification with hydroxynaphthol blue dye, World Journal of Microbiolog y and Biotechnolog y, 33(7), 130 (2017). DOI: https://doi.org/10.1007/s11274-017-2295-5
N.A. Tanner, Y. Zhang, T.C. Evans, Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes, Biotechniques, 58(2), 59-68 (2015). DOI: https://doi.org/10.2144/000114253
M.J. Kellner, J.J. Ross, J. Shnabl, M.P.S. Dekens, R. Heinen, I. Grishkovs-kaya, et al., A rapid, highly sensitive and open-access SARS-CoV-2 detec-tion assay for laboratory and home testing, bioRxlv preprint (2021). Doi: 10.1101/2020.06.23.166397 DOI: https://doi.org/10.1101/2020.06.23.166397
J. Rubinfien, K.D. Atabay, N.M. Nichols, N.A. Tanner, J.A. Pezza, M.M. Gray, et al., Nucleic acid detection aboard the International Space Station by colori-metric loop-mediated isothermal amplification (LAMP), FASEB BioAdvances, 2(3), 160-165 (2020). DOI: https://doi.org/10.1096/fba.2019-00088
M.D. Buck, E.Z. Poirier, A. Cardoso, B. Frederico, J. Canton, S. Barrell, et al., Standard operating procedures for SARS-CoV-2 detection by a clinical diagnostic RT-LAMP assay, medRxiv preprint (2021). Doi: 10.1101/2020.06.29.20142430 DOI: https://doi.org/10.1101/2020.06.29.20142430
L.E. Lamb, S.N. Bartolone, E. Ward, M.B. Chancellor, Rapid detection of novel coronavirus (COVID19) by reverse transcription-loop-mediated isothermal amplification, medRxiv preprint (2021). Doi: 10.1101/2020.02.19.20025155 DOI: https://doi.org/10.2139/ssrn.3539654
Y. Zhang, N. Odiwuor, J. Xiong, L. Sun, R. Nyaruaba, H. Wei, N.A. Tanner, Rapid molecular detection of SARS-CoV-2 (COVID-19) virus RNA using colo-rimetric LAMP, medRxiv preprint (2021). Doi: 10.1101/2020.02.26.20028373 DOI: https://doi.org/10.1101/2020.02.26.20028373
A. Ganguli, A. Mostafa, J. Berger, M. Aydin, F. Sun, S. Ramirez, et al., Rapid isothermal amplification and portable detection system for SARS-CoV-2, Proce-edings of the National Academy of Sciences of the United States of America, 117(37), 22727-22735 (2020). DOI: https://doi.org/10.1073/pnas.2014739117
R. Lu, X. Wu, Z. Wan, Y. Li, L. Zuo, J. Qin, et al.,Development of a novel reverse transcription loop-mediated isothermal amplification method for rapid detec-tion of SARS-CoV-2, Virologica Sinica, 35, 344-347 (2020). DOI: https://doi.org/10.1007/s12250-020-00218-1
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