Detection of dengue viruses using reverse transcription-loop-mediated isothermal amplification

Area of Science:

• Virology
• Molecular Diagnostics
• Public Health

Background:

• Early dengue virus (DENV) detection is vital for patient management and preventing spread.
• Resource-limited settings require sensitive, easy-to-perform diagnostic methods for endemic dengue.

Purpose of the Study:

• To develop and evaluate a novel single-tube reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for DENV detection.
• To assess the clinical applicability of RT-LAMP for diagnosing dengue virus infection in suspected cases.

Main Methods:

• A nine-primer RT-LAMP assay was designed to detect all DENV serotypes and genotypes.
• Sensitivity, specificity, and clinical performance were assessed using 305 serum samples.
• RT-LAMP results were compared against quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and serological assays (IgM/IgG ELISA).

Main Results:

• RT-LAMP detected DENV RNA in 43.3% of confirmed acute infections, comparable to qRT-PCR (46.8%).
• Combining RT-LAMP with IgM/IgG ELISA increased detection to 97.7%, significantly higher than ELISA alone (70.8%).
• The assay demonstrated high concordance with qRT-PCR (κ=0.939), rapid detection (within an hour), and no cross-reactivity with other arboviruses.

Conclusions:

• The developed RT-LAMP assay is sensitive, specific, and user-friendly for dengue diagnosis.
• Integrating RT-LAMP with serological methods enhances the detection rate of acute dengue infections.