Digital LAMP as a point of care assay for HPV detection

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 DESCRIPTION (provided by applicant): We propose to develop a digital loop-mediated-isothermal-amplification (dLAMP), point-of-care (POC), instrument-free, assay for genital human papillomavirus (HPV) testing that can be used as a primary screening method for cervical cancer. Recently, the US Food & Drug Administration approved the Roche Cobas HPV test for stand-alone primary cervical cancer screening. This test, however, requires a sophisticated lab setting and equipment, and thus is not feasible for use as a POC assay, certainly not in a self-test setting, but also not in rural clinical or in resource-poor settings. We have recently developed a novel digital nucleic acid amplification platform based on a self-digitization (SD) microfluidic chip, which partitions an aqueous sample into tens of thousands of nanoliter volumes. Using this SD platform, we have also developed and reported the first digital LAMP assay. Based on these two innovative technologies, we propose to develop a POC HPV detection assay fulfilling the following criteria: (1) It should be as sensitive and specific as existing FDA-approved HPV testing assays for detecting precancerous lesions; (2) It should be instrument-free and require minimal sample preparation, thus offering the flexibility to be done by the patient herself at home; (3) It should provide viral load information, which can be used to improve the specificity of HPV testing and reduce over-treatment; (4) It should be flexible enough to incorporate testing for HPV integration and additional markers to improve specificity; and (5) It should be robust, fast, and affordable. We see application of our proposed POC assay in three different settings: (1) In developed countries, the POC assay can be performed at home and used for primary screening outside the clinical setting, thus maximizing the uptake of screening; (2) In developed countries, the POC assay can be used in the clinic for screening underserved and uninsured populations, allowing for rapid diagnosis and same-visit treatment, thus reducing patient loss to follow-up; (3) In developing countries, the POC assay will allow the clinicians to screen and treat patients at the same clinic visit, again reducing loss of high-risk women through follow-up. Thus, our proposed test will have significant impact on the success of cervical cancer screening in both developed and developing countries.