Development of an Integrated Paper-based Molecular Diagnostic Platform

Point-of-Care testing (POCT) devices have received immense attraction due to its simplicity, ease of use and swiftness. Microfluidic paper-based analytical devices (µPADs) are directly aimed for POC testing. Molecular diagnostics result in a more specific and sensitive assay than immunoassays with much smaller window time to diagnosis. Realizing molecular diagnostics on paper-based devices is a challenge that has been little addressed. A chitosan functionalized paper-based nucleic acid extraction method, isothermal amplification of Chlamydia trachomatis on paper-based devices, and endpoint detection on a nucleic acid lateral flow test strips are presented here. In addition, a proprietary integrated paper-based diagnostic device developed to perform lysate-in-to-answer-out in less than an hour with a limit of detection of 0.2 copies/µL and on going studies to realise a complete system suitable for resource limited settings are also presented. The integrated device is capable of performing affordable, sensitive and specific, and equipment free molecular diagnostic assays in just 6 easy sequential processes.