Low-cost integrated nucleic acid amplification testing platform for cervical cancer screening in low- and middle-income countries - PROJECT SUMMARY/ABSTRACT
An estimated 90% of cervical cancer related deaths take place in low- and middle-income countries (LMICs),
where the burden of disease is highest and there is limited access to effective screening and diagnosis [1]. In
July 2021, the World Health Organization updated their recommended approach for cervical cancer screening
to focus on human papillomavirus (HPV) DNA testing. However, it remains difficult for many women to access
HPV DNA testing due to the necessary costs, high level of infrastructure, and requirement for trained
personnel. Compared to expensive commercial platforms, a variety of promising point-of-care (POC) strategies
have proposed isothermal amplification of DNA as an affordable avenue for testing [2]–[6]; however, there are
important gaps to fully incorporate integrated sample-to-answer platforms that eliminate the risk of workspace
contamination and subsequent false positive results. The objective of this project is to develop a low-cost
(<$3/test, $500/instrument) fully integrated, quantitative HPV DNA test platform that is simple to use for
effective HPV screening at the POC. The Richards-Kortum lab has shown that isothermal amplification with
Recombinase Polymerase Amplification can be performed in paper membranes [7]. More recently, the Posner
group demonstrated the capability to detect discrete fluorescent amplification nucleation sites using a paper
membrane to quantify HIV-1 DNA using a mobile phone-based imager [8]. This proposal builds upon recent
advances to design a two-dimensional paper microfluidic device and low-cost imaging platform for detection of
high risk HPV types, HPV16 and HPV18, the primary causes of cervical cancer. This proposal will accomplish
these goals by pursuing the following three specific aims: 1) Designing a test to accept and process cervical
cytology specimens with pre-loaded lyophilized reagents for amplification of HPV 16/HPV 18 DNA, 2)
Developing a low-cost instrument, including a heater and fluorimeter, to monitor multiplexed real-time
isothermal nucleic acid amplification reactions, and 3) Conducting a proof-of-principle study to assess usability
and efficacy of the test and instrument designed, using banked clinical samples in Houston and within a low
resource setting in Mozambique, Africa. Sensitivity, specificity, and limit of detection analysis will be used to
determine comparability to gold standard technologies. Completing these aims will improve cervical cancer
screening and reduce morbidity and mortality in underserved areas of the world. This project benefits greatly
from co-mentorship from Dr. Rebecca Richards-Kortum and her colleague Kathleen Schmeler, who both have
extensive experience in evaluating new technologies for cervical cancer prevention and translating them to
low-resource settings. The training plan and environment have been specifically tailored, focusing on multi-
disciplinary, hands-on coursework, lab work, and teaching, and translational research aims with deployment
within an LMIC to ensure project success and scalability.
