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 and Rebecca R. Richards-Kortum ()
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Maria J. Barra: Rice University
Alexis F. Wilkinson: Rice University
Ariel E. Ma: Rice University
Karthik Goli: Rice University
Hira Atif: Universidade Eduardo Mondlane (UEM)
Nafissa M.R.B. Osman: Universidade Eduardo Mondlane (UEM)
Cesaltina Lorenzoni: Universidade Eduardo Mondlane (UEM)
Guilhermina Tivir: Centro de Investigação e Treino em Saúde da Polana Caniço (CISPOC)
Eva H. Lathrop: Jhpiego
Philip E. Castle: National Cancer Institute
Ming Guo: The University of Texas MD Anderson Cancer Center
Jane R. Montealegre: The University of Texas MD Anderson Cancer Center
Ellen S. Baker: The University of Texas MD Anderson Cancer Center
Mila P. Salcedo: The University of Texas MD Anderson Cancer Center
Kathleen M. Schmeler: The University of Texas MD Anderson Cancer Center
Rebecca R. Richards-Kortum: Rice University

Nature Communications, 2025, vol. 16, issue 1, 1-12

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-62454-x

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