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.

项目摘要/摘要估计有90％的宫颈癌相关死亡发生在低收入和中等收入国家（LMIC），疾病的燃烧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推荐的宫颈癌筛查方法专注于人乳头瘤病毒（HPV）DNA测试。但是，许多女性仍然很难进入 HPV DNA测试是由于必要的成本，高水平的基础架构以及训练有素的要求人员。与昂贵的商业平台相比，各种有前途的保健（POC）策略提出了DNA作为负担得起的测试途径的等温扩增[2] - [6]；但是，有重要的差距，以充分整合综合样品与答案平台，以消除工作空间的风险污染和随后的假阳性结果。该项目的目的是开发低成本（<$ 3/测试，$ 500/仪器）完全集成的定量HPV DNA测试平台，易于使用有效在POC上进行HPV筛选。 Richards-Kortum实验室表明，等温度放大重组酶聚合酶扩增可以在纸膜中进行[7]。最近，Posner 小组证明了使用纸检测离散荧光扩增核位点的能力使用基于手机的成像仪[8]来量化HIV-1 DNA。该提议建立在最近的设计二维纸微流体设备和低成本成像平台的进步用于检测高风险HPV类型HPV16和HPV18是宫颈癌的主要原因。该建议将完成这些目标通过追求以下三个特定目标：1）设计测试以接受和处理宫颈具有预载的冻干试剂的细胞学标本，用于扩增HPV 16/HPV 18 DNA，2）开发低成本仪器，包括加热器和荧光计，以监视多重实时等温核酸扩增反应，以及3）进行原则证明以评估可用性并易于使用休斯顿的库存临床样品，在低点的情况下设计和仪器设计非洲莫桑比克的资源设定。敏感性，特异性和检测分析的极限将用于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降低世界上服务不足地区的发病率和死亡率。这个项目受益匪浅从丽贝卡·理查兹·科尔图姆（Rebecca Richards-Kortum 在评估预防宫颈癌的新技术方面的丰富经验，并将其转化为低资源设置。培训计划和环境是专门针对的，重点是纪律，动手课程，实验室工作和教学以及翻译研究的目的在LMIC中，以确保项目的成功和可扩展性。