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.

项目摘要/摘要 估计90%的宫颈癌相关死亡发生在低收入和中等收入国家(LMIC)， 疾病负担最重、获得有效筛查和诊断机会有限的地区[1]。在……里面 2021年7月，世界卫生组织更新了他们推荐的宫颈癌筛查方法 将重点放在人类乳头瘤病毒(HPV)DNA检测上。然而，对许多妇女来说，它仍然很难获得 由于必要的成本、高水平的基础设施和对训练有素的人员的要求，HPV DNA检测 人事部。与昂贵的商业平台相比，各种前景看好的医疗点(POC)策略 都建议将DNA等温扩增作为一种负担得起的检测方法[2]-[6]；然而，有 全面整合集成的样本到答案平台以消除工作空间风险的重要差距 污染和随后的假阳性结果。这个项目的目标是开发一种低成本的 (&lt；3美元/检测，500美元/仪器)完全集成的、易于使用的定量HPV DNA检测平台 在POC进行有效的HPV筛查。理查兹-科尔图姆实验室已经证明，等温放大 重组酶聚合酶扩增可以在纸膜中进行[7]。最近，波斯纳 研究小组展示了使用一篇论文检测离散荧光放大成核位点的能力 使用基于移动电话的成像器对HIV-1DNA进行量化的膜。这项建议是建立在最近 二维纸质微流控检测装置及低成本成像平台的设计进展 高危HPV类型，HPV16和HPV18，宫颈癌的主要原因。这项建议将实现 通过追求以下三个具体目标来实现这些目标：1)设计一种接受和处理宫颈病变的测试 用预装冻干试剂扩增HPV16/HPV18DNA的细胞学标本，2) 开发一种低成本的仪器，包括加热器和荧光计，用于实时监测多路传输 等温核酸扩增反应，以及3)进行原理验证研究以评估可用性 以及设计的测试和仪器的有效性，使用休斯顿的银行临床样本并在较低的 非洲莫桑比克的资源环境。将使用灵敏度、特异度和检测限分析 确定与黄金标准技术的可比性。完成这些目标将改善宫颈癌 筛查和减少世界上服务不足地区的发病率和死亡率。这个项目效益很大。 来自丽贝卡·理查兹-科尔图姆博士和她的同事凯瑟琳·施梅勒的共同指导，他们都有 在评估宫颈癌预防新技术并将其转化为 低资源设置。培训计划和环境是专门定制的，重点是多方面的 学科、实践课程、实验室工作和教学，以及翻译研究的目标和部署 在LMIC内确保项目成功和可扩展性。