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

宫颈癌是最可预防和最可治疗的癌症之一，但仍是女性第四常见的癌症。为了在2030年前消除宫颈癌，世卫组织的目标之一是使用高精度检测方法，如HPV检测，对全球70%的35岁至45岁女性进行筛查。在全球每年350,000例与宫颈癌相关的死亡中，大约90%发生在低收入和中等收入国家。这些地区缺乏足够的技术人员、设备齐全的实验室和现有的技术，无法通过市场上现有的测试来实现这一目标。该项目的目标是开发一种护理点诊断工具，以促进近距离患者的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分析，以检测高危HPV并对其进行分型，最终实现自采集拭子。10分钟的样品浓缩和裂解步骤将被优化，而不需要DNA纯化。然后将裂解物用水稀释，并直接送入含有冷冻干燥母料的反应井中，以便在60分钟内进行等温放大。分包商、乔治华盛顿大学传染病分子/微生物学专家珍妮·乔丹教授将在第一阶段结束时验证我们的检测结果。

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.