Simultaneous detection of dengue virus serotypes using a facile gel-free nucleic acid based lateral flow assay
Description
Timely and accurate diagnosis of dengue virus serotypes is crucial for optimal management of patients and control of outbreaks. Diagnostic methods currently available in clinical settings, such as ELISA based tests detect overall DENV infection in patients. In addition, RT-PCR based tests that require specialized equipment and expertise are utilized to detect all four serotypes of DENV. This study focussed on the development of gel free PCR based lateral flow strip assay to detect all four serotypes of DENV on site. The assay uses anti-biotin/streptavidin colloidal gold conjugates with florescent/enzymatic tagged DENV serotype specific antibodies for direct detection on a nitrocellulose membrane using anti-IgG as control line in the DENV infected serum samples. The detection limit of the assay was upto 105 dilutions (10 copies) of initial DENV cDNA. DENV RT-PCR performed to compare the results of the lateral flow assay revealed 100% similarity with the lateral flow results. Results demonstrated 100% sensitivity and specificity for dengue detection and serotype identification.