Abstract
Dengue is a rapidly spreading mosquito-borne viral disease. Early diagnosis is important for clinical screening, medical management, and disease surveillance. The objective of this study was to develop a colorimetric lateral flow biosensor (LFB) for the visual detection of dengue-1 RNA using dextrin-capped gold nanoparticle (AuNP) as label. The detection was based on nucleic acid sandwich-type hybridization among AuNP-labeled DNA reporter probe, dengue-1 target RNA, and dengue-1 specific DNA capture probe immobilized on the nitrocellulose membrane. Positive test generated a red test line on the LFB strip, which enabled visual detection. The optimized biosensor has a cut-off value of 0.01 µM using synthetic dengue-1 target. Proof-of-concept application of the biosensor detected dengue-1 virus in pooled human sera with a cut-off value of 1.2 × 104 pfu/mL. The extracted viral RNA, when coupled with nucleic acid sequence-based amplification (NASBA), was detected on the LFB in 20 min. This study first demonstrates the applicability of dextrin-capped AuNP as label for lateral flow assay. The biosensor being developed provides a promising diagnostic platform for early detection of dengue infection in high-risk resource-limited areas.
Highlights
Dengue is a mosquito-borne disease that affects more than 100 countries in the tropical and sub-tropical regions [1]
Dengue disease is caused by the single-stranded, positive-sense RNA dengue virus (DENV) that occurs in four distinct serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) [3,4]
The principle of detection is based on nucleic acid sandwich reaction on the lateral flow biosensor (LFB) strip
Summary
Dengue is a mosquito-borne disease that affects more than 100 countries in the tropical and sub-tropical regions [1]. Dengue disease is caused by the single-stranded, positive-sense RNA dengue virus (DENV) that occurs in four distinct serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) [3,4]. Primary infection with any of the serotypes can cause dengue fever (DF). Diagnosis of dengue disease based solely on clinical symptoms is difficult and unreliable because initial symptoms of infection are similar to influenza, measles, yellow fever, and other viral infections [7,8]. Current methods used to confirm dengue infection include virus isolation, serological, and molecular techniques [9]. Reverse transcriptase polymerase chain reaction (RT-PCR) remains the gold standard for sensitive molecular and serotyping of dengue virus, but this technique is expensive [10] and rare in endemic areas.
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