Lateral flow immunoassays (LFIAs) were integrated into microfluidic chips and tested to enhance point-of-care testing (POCT), with the aim of improving sensitivity and expanding the range of CRP detection. The microfluidic approach improves upon traditional methods by precisely controlling fluid speed, thus enhancing sensitivity and accuracy in CRP measurements. The microfluidic approach also enables a one-step detection system, eliminating the need for buffer solution steps and reducing the nitrocellulose (NC) pad area to just the detection test line. This approach minimizes the non-specific binding of conjugated antibodies to unwanted areas of the NC pad, eliminating the need to block those areas, which enhances the sensitivity of detection. The gold nanoparticle method detects CRP in the high-sensitivity range of 1–10 μg/mL, which is suitable for chronic disease monitoring. To broaden the CRP detection range, including infection levels beyond 10 μg/mL, fluorescent labels were introduced, extending the measuring range from 1 to 70 μg/mL. Experimental results demonstrate that integrating microfluidic technology significantly enhances operational efficiency by precisely controlling the flow rate and optimizing the mixing efficiency while reducing fabrication resources by eliminating the need for separate pads, making these methods suitable for resource-limited settings. Microfluidics also provides greater control over fluid dynamics compared to traditional LFIA methods, which contributes to enhanced detection sensitivity even with lower sample volumes and no buffer solution, helping to enhance the usability of POCT. These findings highlight the potential to develop accessible, accurate, and cost-effective diagnostic tools essential for timely medical interventions at the POC.