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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要据估计，90% 的宫颈癌相关死亡发生在低收入和中等收入国家 (LMIC)，疾病负担最高且获得有效筛查和诊断的机会有限[1]。 2021年7月，世界卫生组织更新了宫颈癌筛查推荐方法重点关注人乳头瘤病毒 (HPV) DNA 检测然而，许多女性仍然难以获得该检测。由于必要的成本、高水平的基础设施以及训练有素的人员的要求，HPV DNA 检测与昂贵的商业平台相比，各种有前景的即时护理（POC）策略。已经提出将 DNA 等温扩增作为一种经济实惠的测试途径 [2]-[6]；然而，完全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该项目的目标是开发一种低成本的方法。（<3 美元/测试，500 美元/仪器）完全集成的定量 HPV DNA 测试平台，易于使用 Richards-Kortum 实验室已证明，在 POC 中可进行有效的 HPV 筛查。重组酶聚合酶扩增可以在纸膜中进行[7]。研究小组展示了使用纸张检测离散荧光扩增成核位点的能力使用基于移动电话的成像仪来量化 HIV-1 DNA 的膜 [8]。设计二维纸微流体装置和低成本成像平台以检测高危 HPV 类型，HPV16 和 HPV18，是宫颈癌的主要原因。这些目标通过追求以下三个具体目标来实现：1) 设计接受和处理宫颈癌的测试预装冻干试剂的细胞学标本，用于扩增 HPV 16/HPV 18 DNA，2) 开发一种低成本仪器，包括加热器和荧光计，用于实时监测多路复用等温核酸扩增反应，以及 3) 进行原理验证研究以评估可用性使用休斯敦储存的临床样本并在较低的时间内设计测试和仪器的效果和功效非洲莫桑比克的资源设置将用于检测分析的灵敏度、特异性和限度。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。筛查并降低世界服务欠缺地区的发病率和死亡率该项目受益匪浅。来自 Rebecca Richards-Kortum 博士和她的同事 Kathleen Schmeler 的共同指导，他们都在评估宫颈癌预防新技术并将其转化为应用方面拥有丰富的经验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、教学和转化研究目标与部署中低收入国家 (LMIC) 内，以确保项目成功和可扩展性。