One-hour extraction-free loop-mediated isothermal amplification HPV DNA assay for point-of-care testing in Maputo, Mozambique
- Maria J. Barra
- Alexis F. Wilkinson
- Ariel E. Ma
- Karthik Goli
- Hira Atif
- Nafissa M.R.B. Osman
- Cesaltina Lorenzoni
- Guilhermina Tivir
- Eva H. Lathrop
- Philip E. Castle
- Ming Guo
- Jane R. Montealegre
- Ellen S. Baker
- Mila P. Salcedo
- Kathleen M. Schmeler
- Rebecca R. Richards-Kortum
2025-08-07
Human papillomavirus (HPV) is responsible for nearly all cases of cervical cancer. Affordable point-of-care DNA testing is needed for cervical cancer screening in low- and middle-income countries, where most cervical cancer cases occur. HPV DNA testing typically requires complex lab infrastructure and trained personnel. In this work, we develop a loop-mediated isothermal amplification (LAMP)-based HPV DNA test, which targets three of the most oncogenic HPV types (HPV16, HPV18, HPV45) and a cellular control and achieves analytical sensitivity comparable to gold standard methods. Our extraction-free sample preparation strategy permits adding sample lysate directly to the LAMP reaction. We utilize a low-cost benchtop heater/fluorimeter, delivering results in less than one hour. We analytically evaluate our assay with clinical samples in Houston, Texas ( n = 38) and Maputo, Mozambique ( n = 191). Results show 100% and 93% concordance, respectively, with a reference test widely used in low-resource settings. This sensitive and specific four-step assay can potentially expand cervical cancer screening in resource-limited settings.