Low-Cost, Open-Source Colorimetric Platform for Point-Of-Care (POC) Anti-Human Papillomavirus Type 16 (HPV16) Antibody Detection

Human papillomavirus (HPV) is a leading cause of anogenital and head and neck cancers. HPV type 16 (HPV16) is a significant high-risk strain due to its strong association with these malignancies. The burden of HPV-related cancers is particularly severe in low and middle-income countries (LMICs), where access to diagnostic tools is limited. This work introduces a low-cost, point-of-care (POC) colorimetric platform for detection of antibodies against HPV16. The lateral flow immunoassay (LFIA) developed uses immobilized HPV16 CE2 and E7 proteins for antibody capture and colloidal goldconjugated secondary for colorimetric labeling. Quantitative readout is achieved via an imaging device that incorporates open-source electronics and 3D-printed components. Analytical sensitivity experiments determined a limit of detection (LOD) at <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$46 \text{ng} / \text{mL}$</tex>. Clinical evaluation of plasma samples (n <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$=20$</tex>) from head and neck cancer patients <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$(\mathrm{n}=15)$</tex> and healthy individuals <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$(\mathrm{n}=5)$</tex> yielded <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$\mathrm{9 4. 7 \%}$</tex> sensitivity, <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$\mathrm{9 0. 5 \%}$</tex> specificity, and <tex xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">$\mathrm{9 2. 5 \%}$</tex> overall accuracy. This study identified a moderate-to-high prevalence of anti-HPV16 antibodies in head and neck cancer patients, with 66.7 % testing positive. These results suggest that our platform is a viable tool for detection of anti-HPV16 antibodies, particularly in resource-limited settings where cost-effectiveness and accessibility are crucial. Clinical relevance-The reported approach enables screening for HPV-driven oropharengeal and anogenital cancer from a finger stick.