Low-cost integrated nucleic acid amplification testing platform for cervical cancer screening in low- and middle-income countries

用于低收入和中等收入国家宫颈癌筛查的低成本综合核酸扩增检测平台

• 批准号: 10678488
• 负责人: Yajur Naman Maker
• 金额: $ 5.27万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10678488
• 关键词: Acetic Acids; Affect; Africa; Biological Specimen Banks; Buffers; Cell Line; Cellular Phone; Cervical; Cervical Cancer Screening; Cessation of life; Clinical; Collection; Communicable Diseases; Computer software; Country; Cytology; Cytolysis; DNA; DNA amplification; Detection; Development; Diagnosis; Disease; Doctor of Philosophy; Educational process of instructing; Ensure; Environment; Equipment; Fluorescence; Freeze Drying; Genomic DNA; Goals; HIV-1; HPV-High Risk; Heating; Hela Cells; Human Papillomavirus; Human Resources; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus 18; Image; Image Analysis; Improve Access; Infrastructure; Life; Malignant neoplasm of cervix uteri; Membrane; Mentorship; Microfluidic Microchips; Microscope; Monitor; Morbidity - disease rate; Mozambique; Nucleic Acid Amplification Tests; Nucleic Acids; Optics; Pap smear; Paper; Pathology; Patients; Performance; Pilot Projects; Plastics; Polymerase; Preventive measure; Process; Reaction; Reagent; Recommendation; Reporting; Resource-limited setting; Resources; Risk; Sampling; Site; Specificity; Specimen; System; Technology; Testing; Time; Training; Translating; Translational Research; Translations; Tube; Update; Visual; Woman; Work; World Health Organization; burden of illness; cervical cancer prevention; cost; design; detection limit; detection platform; efficacy testing; experience; fluorescence imaging; imager; imaging platform; imaging system; improved; instrument; isothermal amplification; low and middle-income countries; medically underserved; mortality; multidisciplinary; new technology; novel; point of care; recombinase; screening; screening program; success; synthetic construct; two-dimensional; underserved area; underserved community; usability

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.

项目总结/文摘