Point-of-care HPV mRNA test for cervical cancer screening in low-resource settings

在资源匮乏地区进行宫颈癌筛查的护理点 HPV mRNA 检测

• 批准号: 10331882
• 负责人: Rebecca R. Richards-Kortum
• 金额: $ 18.19万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10331882
• 关键词: Address; Benign; Biological Assay; Biological Markers; Biopsy; Cancerous; Cervical; Cervical Cancer Screening; Cessation of life; Climate; Clinical; Clinical Sensitivity; Collaborations; Colposcopy; Complex; Consensus; Country; Cytology; DNA; Detection; Development; Devices; Diagnosis; Diagnostic; Early Diagnosis; Early treatment; Electricity; Epidemiology; Equipment; Future; Goals; HPV E7; HPV-High Risk; Human Papilloma Virus Vaccination; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus 18; Incidence; Industrialization; Infrastructure; Lateral; Lesion; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Measures; Medically Underserved Area; Messenger RNA; Methods; Molecular; Nucleic Acid Amplification Tests; Nucleic Acids; Oncoproteins; Papillomavirus Transforming Protein E6; Patients; Performance; Point-of-Care Systems; Population; Preparation; Prevention program; Process; Production; Publications; RNA; Reagent; Research; Resource-limited setting; Resources; Risk; Sampling; Secondary Prevention; Sensitivity and Specificity; Specificity; System; Technology; Temperature; Test Result; Testing; Texas; Transcript; Translating; Visual; Woman; accurate diagnosis; amplification detection; cancer care; cancer invasiveness; cervical cancer prevention; cost; cost effective; design; detection method; early screening; effectiveness testing; experience; follow-up; improved; innovation; instrument; instrumentation; isothermal amplification; laboratory equipment; low and middle-income countries; medically underserved; mortality; novel; overtreatment; point of care; point of care testing; point-of-care diagnostics; premalignant; prevent; prospective; prototype; public health relevance; randomized trial; screening; screening program; standard of care; stem; viral DNA

Abstract: There is a significant need to improve global access to cervical cancer screening. Globally, approximately 570,000 women are diagnosed with and 311,000 women die from cervical cancer each year. Nearly 90 percent of cervical cancer deaths occur in low- and middle-income countries (LMICs), due mainly to challenges in implementing effective screening programs. Likewise, cervical cancer rates in medically underserved areas in the US remain high. For example, in the Rio Grande Valley of Texas, cervical cancer incidence rates are 55% higher than the US average, and approximately 10% of eligible women are screened. There is broad consensus that high-risk human papillomavirus (hrHPV) testing is the best approach to improve global screening efforts. An HPV biomarker that provides excellent sensitivity and better specificity than DNA is hrHPV mRNA. However, currently available hrHPV mRNA testing remains too complex and costly (e.g. >$45,000 for equipment and roughly $74 per test) for effective implementation into low-resource and medically underserved settings. Advances in isothermal amplification and lateral flow detection offer an opportunity to develop a point-of-care (POC) hrHPV mRNA test that is accurate, affordable, and can be performed in low-resource settings. The goal of this proposal is to combine isothermal amplification and lateral flow detection within an integrated point-of- care device to dramatically lower the cost of hrHPV mRNA testing. We will develop a low-cost, POC hrHPV E7 mRNA test that requires minimal laboratory equipment and performs as well as commercial RNA tests. Isothermal amplification reduces the instrumentation cost and complexity typically associated with nucleic acid amplification requiring only a single-temperature heater. Lateral flow detection integrates sample manipulation processes and wicks all detection reagents past pre-defined test zones, producing a simple, colorimetric readout. Our proposed proof-of-concept test will detect HPV types 16 and 18, the two types responsible for 70% of cervical cancer, and we estimate will cost <$3 per test in low-volume production. Consistent with the exploratory/developmental goals of an R21 proposal, we aim to (1) design and optimize HPV 16 and 18 E7 mRNA amplification assays and lateral flow detection; (2) combine mRNA amplification and detection into a single POC device; (3) evaluate performance of the developed mRNA test using synthetic and clinical samples. We will leverage the expertise of our interdisciplinary team, which includes designing technologies for LMICs, cervical cancer care, HPV diagnostics, and epidemiology, to develop, validate, and translate this novel screening test. We intend to build from this proof-of-concept test to incorporate HPV mRNA detection for types 31,33, 35, 45, 52, and 58 in the future to achieve detection of the HPV types that cause >90% of cervical cancer. Collectively, this research will lead to the development and implementation of a scalable, cost- effective screening test, a critical and necessary step toward the global elimination of cervical cancer.

抽象的： 迫切需要改善全球宫颈癌筛查的可及性。在全球范围内，大约 每年有 570,000 名女性被诊断患有宫颈癌，311,000 名女性死于宫颈癌。接近90% 的宫颈癌死亡发生在低收入和中等收入国家 (LMIC)，主要是由于以下方面的挑战： 实施有效的筛查计划。同样，医疗服务不足地区的宫颈癌发病率 美国仍处于高位。例如，在德克萨斯州里奥格兰德河谷，宫颈癌发病率为 55% 高于美国平均水平，大约 10% 的合格女性接受了筛查。已达成广泛共识 高危人乳头瘤病毒 (hrHPV) 检测是改善全球筛查工作的最佳方法。一个 hrHPV mRNA 是 HPV 生物标志物，比 DNA 具有出色的敏感性和更好的特异性。然而， 目前可用的 hrHPV mRNA 检测仍然过于复杂且昂贵（例如，设备和费用超过 45,000 美元） 每次测试大约 74 美元），以便在资源匮乏和医疗服务不足的环境中有效实施。 等温扩增和侧流检测的进步为开发现场护理提供了机会 (POC) hrHPV mRNA 检测准确、价格实惠，并且可以在资源匮乏的环境中进行。目标 该提案的目的是将等温扩增和侧流检测结合在一个集成的点内 护理设备可大幅降低 hrHPV mRNA 检测的成本。我们将开发一种低成本、POC hrHPV E7 mRNA 测试需要最少的实验室设备，并且执行与商业 RNA 测试相同的效果。 等温扩增降低了通常与核酸相关的仪器成本和复杂性 放大只需要一个单温加热器。侧向层析检测集成了样品操作 处理并吸走所有检测试剂通过预定义的测试区域，产生简单的比色读数。 我们提出的概念验证测试将检测 HPV 16 型和 18 型，这两种类型导致 70% 的宫颈癌 癌症，我们估计小批量生产中每次测试的成本将低于 3 美元。 与 R21 提案的探索/开发目标一致，我们的目标是 (1) 设计和优化 HPV 16和18 E7 mRNA扩增测定和侧流检测； (2)结合mRNA扩增和 检测到单个 POC 设备； (3) 使用合成和评估开发的 mRNA 测试的性能 临床样本。我们将利用跨学科团队的专业知识，其中包括设计 中低收入国家、宫颈癌护理、HPV 诊断和流行病学技术，以开发、验证和 翻译这个新颖的筛选测试。我们打算通过这一概念验证测试来整合 HPV mRNA 未来针对31,33,35,45,52,58型进行检测，实现对引起>90%的HPV型别进行检测 宫颈癌。总的来说，这项研究将导致开发和实施可扩展的、成本低廉的 有效的筛查测试是全球消除宫颈癌的关键且必要的一步。