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Diagn Microbiol Infect Dis. 2015 Sep;83(1):30-6. doi: 10.1016/j.diagmicrobio.2015.05.004. Epub 2015 May 15.

Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus.

Author information

1
Naval Medical Research Center, Silver Spring, MD 20910-7500, USA.
2
Naval Medical Research Center, Silver Spring, MD 20910-7500, USA. Electronic address: subhamoy.pal.ctr@mail.mil.
3
National Chung Hsing University, Taichung, Taiwan.
4
US Naval Medical Research Unit-6, Iquitos, Peru.

Abstract

During dengue outbreaks, acute diagnosis at the patient's point of need followed by appropriate supportive therapy reduces morbidity and mortality. To facilitate needed diagnosis, we developed and optimized a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay that detects all 4 serotypes of dengue virus (DENV). We used a quencher to reduce nonspecific amplification. The assay does not require expensive thermocyclers, utilizing a simple water bath to maintain the reaction at 63 °C. Results can be visualized using UV fluorescence, handheld readers, or lateral flow immunochromatographic tests. We report a sensitivity of 86.3% (95% confidence interval [CI], 72.7-94.8%) and specificity of 93.0% (95% CI, 83.0-98.1%) using a panel of clinical specimens characterized by DENV quantitative reverse transcription-polymerase chain reaction. This pan-serotype DENV RT-LAMP can be adapted to field-expedient formats where it can provide actionable diagnosis near the patient's point of need.

KEYWORDS:

Dengue; Loop-mediated isothermal amplification; RT-LAMP

[Indexed for MEDLINE]

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