Paper-based microfluidics is a subarea of microfluidics which is recently used in various applications from diagnostics to environmental monitoring, and to food safety. In such microfluidic systems, a test platform is formed from a paper substrate instead of silicon and polymers, such as poly-dimethylsiloxane, poly-methyl methacrylate, and etc. The main goal of this thesis is the development and fabrication of a paper-based microfluidic device (μPAD), which could be used in point-of-care (POC) applications. The characterizations of μPADs, which were fabricated via laser ablation methodology, were performed in terms of their surface and barrier characteristics, and liquid sample flows within μPADs. Depending on the characterization, nine different fabrication parameters, 10P40S (10%Power & 40%Speed), 10P60S, 20P90S, 30P50S, 30P100S, 40P80S, 40P100S, 70P80S, and 70P100S, were identified as optimized fabrication parameters. Also, two designed models of μPADs, 1S4T-Type2 and 1S4T-Type3, were selected to be used in the detection of BSA and recombinant Hepatitis C Virus (HCV) protein. The BSA and HCV (1 mg/ml) in PBS solution were successfully detected via naked eye depending on the colorimetric sensing through micro-paper enzyme linked immunosorbent assay (μP-ELISA) protocol. Moreover, the limit of detection (LoD) values for HCV were determined in 1S4T-Type2 μPAD as 1.000, 0.883, and 0.796 ng/ml when the detection was performed via naked eye, smart-phone, and bright-field microscope, respectively. Also, the easily-disposable 1S4T-Type2 μPAD provided 14 times faster and 45 times cheaper detection of HCV compared to conventional ELISA techniques. Consequently, the developed 1S4T-Type2 μPAD presented low-cost, easy-to-use, and rapid detection of HCV as POC devices.