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:
  • 金额:
    $ 5.27万
  • 依托单位:
  • 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

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.
项目总结/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,获得有效筛查和诊断的机会有限[1]。在 2021年7月,世界卫生组织更新了其推荐的宫颈癌筛查方法 关注人类乳头瘤病毒(HPV)DNA检测。然而,许多妇女仍然难以获得 HPV DNA检测由于必要的成本,高水平的基础设施,以及对培训的要求, 人员的与昂贵的商业平台相比,各种有前途的床旁(POC)策略 已经提出了DNA等温扩增作为一种负担得起的测试途径[2]-[6];然而, 在充分纳入消除工作空间风险的综合抽样到回答平台方面存在重大差距 污染和随后的假阳性结果。该项目的目标是开发一种低成本的 (<3美元/检测,500美元/仪器)完全集成的HPV DNA定量检测平台,使用简单, 在POC进行有效的HPV筛查。Richards-Kortum实验室已经证明, 聚合酶扩增可以在纸膜中进行[7]。最近,波斯纳 小组证明了使用纸张检测离散荧光扩增成核位点的能力 使用基于移动的手机成像仪对膜进行HIV-1 DNA定量[8]。该提案建立在最近 二维纸微流控装置和低成本成像平台的设计进展, 高危型HPV,HPV 16和HPV 18,宫颈癌的主要原因。这项提案将实现 通过追求以下三个具体目标来实现这些目标:1)设计一个测试来接受和处理宫颈癌 用于扩增HPV 16/HPV 18 DNA的预装冻干试剂的细胞学标本,2) 开发了一种低成本的仪器,包括加热器和荧光计,以实时监测多路复用 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 使用休斯顿的库存临床样本和低浓度范围内, 非洲莫桑比克的资源环境。灵敏度、特异性和检测限分析将用于 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 筛查和减少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 Rebecca Richards-Kortum博士和她的同事Kathleen Schmeler的共同指导, 在评估预防宫颈癌的新技术并将其转化为 低资源环境。培训计划和环境经过专门定制,侧重于多方面, 学科,实践课程,实验室工作,教学和翻译研究的目标与部署 确保项目成功和可扩展性。

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