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

子宫颈癌是最容易预防和治疗的癌症之一，但仍然是第四大最常见的女性癌症。为了到2030年消除宫颈癌，世卫组织的目标之一是使用HPV检测等高精度检测对全球70%的35至45岁女性进行筛查。全球每年35万宫颈癌相关死亡中，约90%发生在低收入和中等收入国家。这些地区没有足够的熟练人员、设备齐全的实验室和技术来实现这一目标，目前市场上有检测。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、空间复用、实时等温扩增的检测方法，通过最终的自收集拭子来检测高危HPV并进行基因分型。优化10分钟的样品浓缩和裂解步骤，无需DNA纯化。然后将裂解物用水稀释，并直接输送到含有冻干主混合物的反应井中，在< 60分钟内进行等温扩增。分包商，乔治华盛顿大学传染病分子/微生物学专家Jeanne Jordan教授将在第一阶段结束时验证我们的分析。

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.