Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification

PLoS One. 2015 Sep 18;10(9):e0138694. doi: 10.1371/journal.pone.0138694. eCollection 2015.

Abstract

Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Case-Control Studies
  • Colorimetry / methods*
  • Dengue / blood
  • Dengue / diagnosis*
  • Humans
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity

Grants and funding

The funding for this study was changed to High Impact Research Grant UM-MOHE UM.C/625/1/HIR/MOHE/CHAN/14/3 from the Ministry of Higher Education Malaysia and University Malaya Research Grant LR001-2011D.