Molecular diagnostics of dengue by reverse transcription-loop mediated isothermal amplification (RT-LAMP) in disposable polyester-toner microdevices
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Data
2019-05-14
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Resumo
Dengue is one of the most prevalent infectious tropical diseases in the world, with high
incidence in over 100 countries. Rapid and reliable diagnosis of dengue is of great importance
for public health. To simplify molecular diagnostics, isothermal amplification techniques have
recently emerged as an alternative to conventional methods of deoxyribonucleic acid (DNA)
amplification. Here, we developed a one-step method for dengue virus detection from real sample
based on RT-LAMP (reverse transcription-loop mediated isothermal amplification) in a disposable
microdevice. The reaction was thermally controlled with a thermoblock for 15 min at 72 °C. At
the end of the incubation time, we either removed the solution for detection of fragments by gel
electrophoresis or added DNA intercalator for visual detection on-chip. Our results demonstrated
that it is possible to detect dengue virus through RT-LAMP directly from a serum sample, without
previous ribonucleic acid (RNA) extraction. The success of RT-LAMP was confirmed in reactions
initiated with 0.8 fg μL−1 of RNA, which represents 200 copies of RNA per µL. RT-LAMP in
a polyester-toner (PeT) microdevice is a simple and inexpensive method that allows for rapid
detection of dengue virus with high reliability and great potential for point-of-care applications.
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RT-LAMP, PeT microdevice, Dengue virus, Molecular diagnosis, Point-of-care
Citação
MENDES, Geovana M. et al. Molecular diagnostics of dengue by reverse transcription-loop mediated isothermal amplification (RT-LAMP) in disposable polyester-toner microdevices. Journal of the Brazilian Chemical Society, Campinas, v. 30, n. 9, p. 1841-1849, 2019. DOI: 10.21577/0103-5053.20190092. Disponível em: https://jbcs.sbq.org.br/default.asp?ed=289. Acesso em: 13 dez. 2023.