One-hour extraction-free loop-mediated isothermal amplification HPV DNA assay for point-of-care testing in Maputo, Mozambique

Résumé

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.

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