Colorimetric On-Chip Detection of Human Papillomavirus for Point-Of-Care Settings

Point-of-care diagnostics have evolved over the past decade to be a feasible alternative to traditional qPCR testing. Isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) detection have become leading POC diagnostic tools and can be used for cancer-causing persistent viral infections such as high-risk human papillomavirus (HPV). Here, we have developed a colorimetric LAMP assay targeting HPV 16 and 18 L 1 regions for ease of use and instrument-free detection. The diagnostic system comprises of a transparent, 3D-printed microfluidic chip with four reaction wells for simultaneous testing and controls. Validation with spiked synthetic templates demonstrated the LAMP assay's sensitivity, detecting as few as 10-100 copies of HPV DNA per reaction. The limit of detection (LOD) was confirmed through serial dilutions of HPV 16 DNA from 1 to 10<sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">7</sup> copies. Integration with a syringe and luer lock inlet port facilitated sample treatment, and the assay was tested for both colorimetric and fluorescence detection. Future developments will focus on field testing and clinical validation, inclusion of additional HPV types, and further optimization for routine cervical cancer screening. This POC system offers a promising, rapid, and accessible method for early HPV detection, particularly in resource-limited settings. Clinical relevance-We present an approach to screening for HPV-driven cervical cancer for low resource settings. The colorimetric readout along with the isothermal reaction make the approach reduce the need for expensive, high-precision instrumentation.