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Submitted: 05 July 2013 Modified: 18 January 2017

Herdin Record #: NCR-RITM-13070512545819

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

Researchers

NameRole
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

Publication Information

1.
Publication Type:
Journal
Publication Sub Type:
Journal Article, Original
Title:
Journal of Clinical Microbiology
Frequency:
Annual
Publication Date:
June 2005
Volume:
43
Issue:
6
Page(s):
2895–2903
Publisher:
American Society for Microbiology

Abstract

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.


Physical Location

LocationCall NumberAvailable FormatAvailability
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)