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
- 来源:
- 关键词:Acetic AcidsAffectAfricaBiological Specimen BanksBuffersCell LineCellular PhoneCervicalCervical Cancer ScreeningCessation of lifeClinicalCollectionCommunicable DiseasesComputer softwareCountryCytologyCytolysisDNADNA amplificationDetectionDevelopmentDiagnosisDiseaseDoctor of PhilosophyEducational process of instructingEnsureEnvironmentEquipmentFluorescenceFreeze DryingGenomic DNAGoalsHIV-1HPV-High RiskHeatingHela CellsHuman PapillomavirusHuman ResourcesHuman papilloma virus infectionHuman papillomavirus 16Human papillomavirus 18ImageImage AnalysisImprove AccessInfrastructureLifeMalignant neoplasm of cervix uteriMembraneMentorshipMicrofluidic MicrochipsMicroscopeMonitorMorbidity - disease rateMozambiqueNucleic Acid Amplification TestsNucleic AcidsOpticsPap smearPaperPathologyPatientsPerformancePilot ProjectsPlasticsPolymerasePreventive measureProcessReactionReagentRecommendationReportingResource-limited settingResourcesRiskSamplingSiteSpecificitySpecimenSystemTechnologyTestingTimeTrainingTranslatingTranslational ResearchTranslationsTubeUpdateVisualWomanWorkWorld Health Organizationburden of illnesscervical cancer preventioncostdesigndetection limitdetection platformefficacy testingexperiencefluorescence imagingimagerimaging platformimaging systemimprovedinstrumentisothermal amplificationlow and middle-income countriesmedically underservedmortalitymultidisciplinarynew technologynovelpoint of carerecombinasescreeningscreening programsuccesssynthetic constructtwo-dimensionalunderserved areaunderserved communityusability
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]。在
2021年7月,世界卫生组织更新了宫颈癌筛查推荐方法
专注于人乳头瘤病毒 (HPV) DNA 检测。然而,许多女性仍然很难获得
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
人员。与昂贵的商业平台相比,各种有前途的即时护理(POC)策略
已提出 DNA 等温扩增作为一种经济实惠的测试途径 [2]–[6];然而,有
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
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
(<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
在 POC 进行有效的 HPV 筛查。 Richards-Kortum 实验室表明,等温扩增
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纸张检测离散荧光扩增成核位点的能力
使用基于手机的成像仪对 HIV-1 DNA 进行定量 [8]。该提案建立在最近
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。该提案将实现
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配置。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
在世界服务欠缺地区进行筛查并降低发病率和死亡率。这个项目受益匪浅
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技术方面拥有丰富的经验
低资源设置。培训计划和环境是专门定制的,重点关注多方面
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。
项目成果
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