1. An integrated isothermal nucleic acid amplification test to detect HPV16 and HPV18 DNA in resource-limited settings.
- Author
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Kundrod, Kathryn A., Barra, Maria, Wilkinson, Alexis, Smith, Chelsey A., Natoli, Mary E., Chang, Megan M., Coole, Jackson B., Santhanaraj, Akshaya, Lorenzoni, Cesaltina, Mavume, Celda, Atif, Hira, Montealegre, Jane Richards, Scheurer, Michael E., Castle, Philip E., Schmeler, Kathleen M., and Richards-Kortum, Rebecca R.
- Subjects
NUCLEIC acid amplification techniques ,RESOURCE-limited settings ,HUMAN papillomavirus ,GENE amplification ,EARLY detection of cancer ,POLYMERASES - Abstract
High-risk human papillomavirus (HPV) DNA testing is widely acknowledged as the most sensitive cervical cancer screening method but has limited availability in resource-limited settings, where the burden of cervical cancer is highest. Recently, HPV DNA tests have been developed for use in resource-limited settings, but they remain too costly for widespread use and require instruments that are often limited to centralized laboratories. To help meet the global need for low-cost cervical cancer screening, we developed a prototype, sample-to-answer, point-of-care test for HPV16 and HPV18 DNA. Our test relies on isothermal DNA amplification and lateral flow detection, two technologies that reduce the need for complex instrumentation. We integrated all test components into a low-cost, manufacturable platform, and performance of the integrated test was evaluated with synthetic samples, provider-collected clinical samples in a high-resource setting in the United States, and self-collected clinical samples in a low-resource setting in Mozambique. We demonstrated a clinically relevant limit of detection of 1000 HPV16 or HPV18 DNA copies per test. The test requires six user steps, yields results in 45 min, and can be performed using a benchtop instrument and minicentrifuge by minimally trained personnel. The projected per-test cost is <$5, and the projected instrumentation cost is <$1000. These results show the feasibility of a sample-to-answer, point-of-care HPV DNA test. With the inclusion of other HPV types, this test has the potential to fill a critical gap for decentralized and globally accessible cervical cancer screening. Editor's summary: DNA-based screening methods for high-risk human papillomavirus (HPV) have proven to be highly effective for early cervical cancer detection but remain too expensive and difficult to use in resource-limited settings. Kundrod et al. developed a prototype, sample-to-answer, point-of-care test for detecting HPV16 and HPV18. This test uses isothermal DNA amplification and lateral flow detection with low-cost components that can detect HPV DNA at a clinically relevant limit of detection. This test was successfully demonstrated in a high-resource setting in the United States and in a low-resource setting in Mozambique, and it could be performed by minimally trained personnel at an estimated cost of <$5 per test. These results suggest that a sample-to-answer, point-of-care HPV DNA test is viable for low-resource settings and could improve access to cervical cancer screening. —Christiana Fogg [ABSTRACT FROM AUTHOR]
- Published
- 2023
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