Bioinformatics-based identification and preliminary clinical evaluation of molecular biomarkers for LAMP-based diagnosis of lymphatic filariasis

Lymphatic filariasis (LF), predominantly caused by Wuchereria bancrofti , requires rapid and accurate diagnostics to support elimination programs, particularly in resource-limited settings. The present study exemplified the development and laboratory validation of a loop-mediated isothermal amplification (LAMP) assay for detecting W. bancrofti DNA. We identified five W. bancrofti -specific gene markers by using a bioinformatics-driven target selection approach. The assay achieved a detection limit of picograms in spiked blood samples, thereby minimizing false-positive rates and showing no cross-reactivity with DNA from Brugia malayi , Brugia pahangi , Dirofilaria immitis and other bacterial and fungal pathogens. A colorimetric readout using phenol red enabled simple visual interpretation of endpoint results, eliminating the need for specialized instrumentation, while a ready-to-use master mix enhanced usability and consistency. The LAMP assay was validated by spiking W. bancrofti DNA into human blood to mimic patient samples. Additionally, a preliminary clinical evaluation was performed by testing eight LF patient samples. The LAMP assay offers increased sensitivity and specificity, with a 30–60 min turnaround time from sample collection to result. LAMP assay offers a visual readout-based point-of-care test for monitoring and management of W. bancrofti infection in endemic and resource-limited settings.