A RAPID ISOTHERMAL AMPLIFICATION CHIP FOR HPV DETECTION AND GENOTYPING IN LOW-RESOURCE SETTINGS

用于低资源环境下 HPV 检测和基因分型的快速等温扩增芯片

• 批准号: 10707836
• 负责人: SEASON WONG
• 金额: $ 39万
• 依托单位: AI BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2023-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10707836
• 关键词: Area; Biological Assay; Businesses; Cessation of life; Characteristics; Collection; Communicable Diseases; Cytolysis; DNA purification; Detection; Diagnostic tests; Evaluation; Freeze Drying; Genotype; HPV-High Risk; Human Papillomavirus; Human Resources; Jordan; Laboratories; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Microbiology; Molecular; Patients; Performance; Phase; Process; Protocols documentation; Reaction; Resource-limited setting; Sampling; Sensitivity and Specificity; Signal Transduction; Swab; Technology; Testing; Time; Universities; Washington; Water; Woman; base; clinically relevant; cross reactivity; design; detection limit; diagnostic tool; isothermal amplification; low and middle-income countries; multiplex assay; point-of-care diagnostics; professor; stability testing

Cervical cancer is one of the most preventable and treatable forms of cancer, but still the fourth most common cancer in women. To eliminate cervical cancer by 2030, one of the WHO’s targets is to screen 70% of 35 to 45-year old women globally using a high-precision test, such as an HPV test. Approximately 90% of the globe’s 350,000 annual cervical cancer related deaths occur in low- and middle-income countries. These areas are insufficiently equipped with the skilled personnel, well-equipped labs, and technologies available to reach this target with tests currently available on the market. The objective of this project is to develop a point-of-care diagnostic tool to advance near-patient HPV detection and genotyping. We will validate a chip- based, spatial-multiplexed, real-time isothermal amplification assay to detect and genotype high-risk HPV with eventual self-collected swab implementation. A 10-minute process of sample concentration and lysing steps without DNA purification will be optimized. The lysate will then be diluted with water and delivered directly into reaction wells that contain lyophilized master mix for the isothermal amplification in < 60 minutes. Subcontractor, Professor Jeanne Jordan, a molecular/microbiology expert for infectious diseases at George Washington University will validate our assay at the end of Phase I.

宫颈癌是最可预防和治疗的癌症之一，但仍然是妇女第四大常见癌症。为了到2030年消除宫颈癌，世卫组织的目标之一是使用高精度检测（如HPV检测）对全球70%的35至45岁女性进行筛查。地球仪每年350，000例宫颈癌相关死亡中约90%发生在低收入和中等收入国家。这些地区没有足够的技术人员、设备齐全的实验室和可用的技术，无法通过目前市场上的测试达到这一目标。该项目的目标是开发一种即时诊断工具，以推进近患者HPV检测和基因分型。我们将验证一种基于芯片的、空间多路复用的、实时等温扩增检测方法，以检测高危型HPV并进行基因分型，最终实现自我采集拭子。将优化10分钟的样品浓缩和裂解步骤过程，无需DNA纯化。 然后将裂解物用水稀释，并直接递送到含有冻干主混合物的反应威尔斯孔中，用于在< 60分钟内进行等温扩增。此外，乔治华盛顿大学传染病分子/微生物学专家Jeanne Jordan教授将在I期结束时验证我们的检测方法。