Keywords
Amplificación isotérmica
Malaria
Plasmodium
Diagnóstico
Malaria
Plasmodium
Diagnóstico
How to Cite
Arroyo A, M. I., Morales L, G. P., Sosa V, P. A., Carmona-Fonseca, J., & Maestre B, A. (2020). Amplificación isotérmica de ácidos nucleicos tipo LAMP para la detección de Plasmodium: nueva técnica diagnóstica. Médicas UIS, 21(3), 158–175. Retrieved from https://revistas.uis.edu.co/index.php/revistamedicasuis/article/view/12314
Abstract
Recientemente se introdujo en el diagnóstico de malaria una técnica llamada amplifi cación isotérmica de ácidos nucleicos, que usa el material genético plasmodial. Este escrito revisa la información disponible sobre amplifi cación isotérmica de ácidos nucleicos, especialmente en el campo del paludismo. Metodología: se revisaron las bases electrónicas Lilacs, Scielo, PubMed (Medline) y Ovid. Resultados: solo se encontraron tres referencias sobre amplifi cación isotérmica de ácidos nucleicos y malaria pero hubo abundante información sobre amplifi cación isotérmica de ácidos nucleicos en otras infecciones y campos de la medicina, en especial la infectología. La reacción de amplifi cación isotérmica de ácidos nucleicos requiere de una ADN polimerasa con actividad de desplazamiento de cadena y cuatro cebadores especialmente diseñados para reconocer seis secuencias distintas. Esto garantiza alta especifi cidad para la amplifi cación. Varias alternativas de amplifi cación isotérmica de ácidos nucleicos se han desarrollado para la identifi cación de virus, bacterias, micoplasmas, protozoos, hongos y levaduras. Ventajas de amplifi cación isotérmica de ácidos nucleicos son capacidad de amplifi car ácidos nucleicos bajo condiciones isotérmicas (60-65 ºC); posibilidad de lectura y semicuantifi cación a simple vista y de cuantifi cación con un turbidímetro; altas sensibilidad y especifi cidad y, en general, capacidad diagnóstica; rapidez, bajo costo y facilidad de aplicación; tolera los componentes de los medios de cultivo y las sustancias biológicas. Entre las desventajas están que la baja concentración de ADN molde disminuye la efi cacia del reconocimiento de los ebadores; la observación de la turbidez fue menos sensible que la visualización de los productos en gel de agarosa teñidos con bromuro de etidio y es crítica la revención de la contaminación. (MÉD.UIS. 2008;21(3):158-75).
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13. Tham J, Lee S, Tan T, Ting R, Kara U. Detection and species
determination of malaria parasites by PCR: comparison with
microscopy and with ParaSight-F and ICT Malaria Pf tests in
a clinical environment. J Clin Microbiol. 1999;37:1269–73.
14. Boonma P, Christensen P, Suwanarusk R, Price R, Russell B,
Lek-Uthai U. Comparison of three molecular methods for the
detection and speciation of Plasmodium vivax and
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15. Gama BE, Silva-Pires F, Lopes M, Cardoso M, Britto C, Torres
K, et al. Real-time PCR versus conventional PCR for malaria
parasite detection in low-grade parasitemia. Exp Parasitol.
2007;16:427-32.
16. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe
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DNA. Nucleic Acids Res. 2000;28:E63.
17. Notomi T. [Loop-mediated isothermal amplification] [artículo
en japonés; resumen en inglés]. Nippon Rinsho. 2007;65:957-
61.
18. Tomita N, Mori Y, Kanda H, Notomi T. Loop-mediated
isothermal amplification (LAMP) of gene sequences and
simple visual detection of products. Nature Protocols.
2008;3:877-82.
19. Snounou G. Rapid, sensitive and cheap molecular diagnosis
of malaria: is microscopy on the way out? Future Microbiology.
2007;2:477-80.
20. Imai M, Ninomiya A, Minekawa H, Notomi T, Ishizaki T,
Tashiro M, et al. Development of H5-RT-LAMP (loop-mediated
isothermal amplification) system for rapid diagnosis of H5
avian influenza virus infection. Vaccine. 2006;24:6679-82
21. Kurosaki Y, Takada A, Ebihara H, Grolla A, Kamo N, Feldmann
H, et al. Rapid and simple detection of Ebola virus by reverse
transcription-loop-mediated isothermal amplification. J Virol
Methods. 2007;141:78-83.
22. Kuhara T, Yoshikawa T, Ihira M, Watanabe D, Tamada Y,
Katano H, et al. Rapid detection of human herpesvirus 8
DNA using loop-mediated isothermal amplification. J Virol
Methods. 2007;144:79-85.
23. Mori N, Motegi Y, Shimamura Y, Ezaki T, Natsumeda T,
Yonekawa T, et al. Development of a new method for
diagnosis of rubella virus infection by reverse
transcription-loop-mediated isothermal amplification. J
Clin Microbiol. 2006;44:3268-73.
24. Okafuji T, Yoshida N, Fujino M, Motegi Y, Ihara T, Ota Y,
et al. Rapid diagnostic method for detection of mumps
virus genome by loop-mediated isothermal amplification.
J Clin Microbiol. 2005;43:1625-31.
25. Fujino M, Yoshida N, Yamaguchi S, Hosaka N, Ota Y,
Notomi T, et al. A simple method for the detection of
measles virus genome by loop-mediated isothermal
amplification (LAMP). J Med Virol. 2005;76:406-13.
26. Kaneko H, Iida T, Aoki K, Ohno S, Suzutani T. Sensitive
and rapid detection of herpes simplex virus and varicellazoster virus DNA by loop-mediated isothermal
amplification. J Clin Microbiol. 2005;43:3290-6.
27. Enomoto Y, Yoshikawa T, Ihira M, Akimoto S, Miyake F,
Usui C, et al. Rapid diagnosis of herpes simplex virus
infection by a loop-mediated isothermal amplification
method. J Clin Microbiol. 2005;43:951-5.
28. Suzuki R, Yoshikawa T, Ihira M, Enomoto Y, Inagaki S,
Matsumoto K, et al. Development of the loop-mediated
isothermal amplification method for rapid detection of
cytomegalovirus DNA. J Virol Methods. 2006;132:216-21.
29. Iwata S, Shibata Y, Kawada J, Hara S, Nishiyama Y,
Morishima T, et al. Rapid detection of Epstein-Barr virus
DNA by loop-mediated isothermal amplification method.
J Clin Virol. 2006;37:128-33.
30. Yamada Y, Itoh M, Yoshida M. Sensitive and rapid diagnosis
of human parvovirus B19 infection by loop-mediated
isothermal amplification. Br J Dermatol. 2006;155:50-5.
31. Wakabayashi T, Yamashita R, Kakita T, Kakita M, Oshika T.
Rapid and sensitive diagnosis of adenoviral
keratoconjunctivitis by loop-mediated isothermal
amplification (LAMP) method. Curr Eye Res. 2004;28(6):445-
50.
32. Parida M, Horioke K, Ishida H, Dash P, Saxena P, Jana A, et
al. Rapid detection and differentiation of dengue virus
serotypes by a real-time reverse transcription-loopmediated isothermal amplification assay. J Clin Microbiol.
2005;43:2895-903.
33. Parida M, Santhosh S, Dash P, Tripathi N, Saxena P, Ambuj
S, et al. Development and evaluation of reverse
transcription-loop-mediated isothermal amplification
assay for rapid and real-time detection of Japanese
encephalitis virus. J Clin Microbiol. 2006;44:4172-8.
34. Poon L, Leung C, Chan K, Lee J, Yuen K, Guan Y, et al.
Detection of human influenza A viruses by loop-mediated
isothermal amplification. J Clin Microbiol. 2005;43:427-
30. 35. Ito M, Watanabe M, Nakagawa N, Ihara T, Okuno Y. Rapid
detection and typing of influenza A and B by loopmediated isothermal amplification: comparison with
immunochromatography and virus isolation. J Virol
Methods. J Virol Methods. 2006;135:272-5.
36. Yoneyama T, Kiyohara T, Shimasaki N, Kobayashi G, Ota
Y, Notomi T, et al. Rapid and real-time detection of
hepatitis A virus by reverse transcription loop-mediated
isothermal amplification assay. J Virol Methods.
2007;145:162-8.
37. Minami M, Ohta M, Ohkura T, Ando T, Torii K, Hasegawa
T, et al. Use of a combination of brushing technique and
the loop-mediated isothermal amplification method as a
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