There is a significant need to improve global access to cervical cancer screening. Globally, approximately 570,000 women are diagnosed with and 311,000 women die from cervical cancer each year. Nearly 90 percent of cervical cancer deaths occur in low- and middle-income countries (LMICs), due mainly to challenges in implementing effective screening programs. Likewise, cervical cancer rates in medically underserved areas in the US remain high. For example, in the Rio Grande Valley of Texas, cervical cancer incidence rates are 55% higher than the US average, and approximately 10% of eligible women are screened. There is broad consensus that high-risk human papillomavirus (hrHPV) testing is the best approach to improve global screening efforts. An HPV biomarker that provides excellent sensitivity and better specificity than DNA is hrHPV mRNA. However, currently available hrHPV mRNA testing remains too complex and costly (e.g. >$45,000 for equipment and roughly $74 per test) for effective implementation into low-resource and medically underserved settings. Advances in isothermal amplification and lateral flow detection offer an opportunity to develop a point-of-care (POC) hrHPV mRNA test that is accurate, affordable, and can be performed in low-resource settings. The goal of this proposal is to combine isothermal amplification and lateral flow detection within an integrated point-ofcare device to dramatically lower the cost of hrHPV mRNA testing. We will develop a low-cost, POC hrHPV E7 mRNA test that requires minimal laboratory equipment and performs as well as commercial RNA tests. Isothermal amplification reduces the instrumentation cost and complexity typically associated with nucleic acid amplification requiring only a single-temperature heater. Lateral flow detection integrates sample manipulation processes and wicks all detection reagents past pre-defined test zones, producing a simple, colorimetric readout. Our proposed proof-of-concept test will detect HPV types 16 and 18, the two types responsible for 70% of cervical cancer, and we estimate will cost <$3 per test in low-volume production. Consistent with the exploratory/developmental goals of an R21 proposal, we aim to (1) design and optimize HPV 16 and 18 E7 mRNA amplification assays and lateral flow detection; (2) combine mRNA amplification and detection into a single POC device; (3) evaluate performance of the developed mRNA test using synthetic and clinical samples. We will leverage the expertise of our interdisciplinary team, which includes designing technologies for LMICs, cervical cancer care, HPV diagnostics, and epidemiology, to develop, validate, and translate this novel screening test. We intend to build from this proof-of-concept test to incorporate HPV mRNA detection for types 31,33, 35, 45, 52, and 58 in the future to achieve detection of the HPV types that cause >90% of cervical cancer. Collectively, this research will lead to the development and implementation of a scalable, costeffective screening test, a critical and necessary step toward the global elimination of cervical cancer.