Herdin Record #: NCR-RITM-13070512545819

Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription–loop-mediated isothermal amplification assay

Name	Role
1Manmohan Parida	Author
2Kouhei Horioke	Author
3Hiroyuki Ishida	Author
4Paban Kumar Dash	Author
5Parag Saxena	Author
6Asha Mukul Jana	Author
7Mohammed Alimul Islam	Author
8Shingo Inoue	Author
9Norimitsu Hosaka	Author
10Kouichi Morita	Author

The development and validation of a one-step, real-time, and quantitative dengue virus serotype-specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay targeting the 3
noncoding region for the rapid detection and differentiation of dengue virus serotypes are reported. The RT-LAMP assay is very simple and rapid, wherein the amplification can be obtained in 30 min under isothermal conditions at
63°C by employing a set of four serotype-specific primer mixtures through real-time monitoring in an inexpensive turbidimeter. The evaluation of the RT-LAMP assay for use for clinical diagnosis with a limited number of patient serum samples, confirmed to be infected with each serotype, revealed a higher sensitivity by picking up 100% samples as positive, whereas 87% and 81% of the samples were positive by reverse transcription-PCR and virus isolation, respectively. The sensitivity and specificity of the RT-LAMP assay for the detection of viral RNA in patient serum samples with reference to virus isolation were 100% and 93%, respectively. The optimal assay conditions with zero background and no cross-reaction with other closely related members of the Flavivirusfamily (Japanese encephalitis, West Nile, and St. Louis encephalitis viruses) as well as within the four serotypes of dengue virus were established. None of the serum samples from healthy individuals screened in this study showed any cross-reaction with the four dengue virus serotype-speci?c RT-LAMP assay primers. These findings demonstrate that RT-LAMP assay has the potential clinical application for detection and differentiation of dengue virus serotypes, especially in developing countries.

Location	Call Number	Available Format	Availability
University of the Philippines		Fulltext	Print Format
U.S. National Library of Medicine: PubMed/Medline		Abstract	External Link (View)
U.S. National Library of Medicine: PubMed/Medline		Fulltext	pdf (Request Document)