Low-Cost CRISPR-on-Paper for Cervical Cancer Screening at the Point of Care

用于宫颈癌护理点筛查的低成本纸上 CRISPR

• 批准号: 10415457
• 负责人: Changchun Liu
• 金额: $ 54.73万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10415457
• 关键词: 2019-nCoV; Acetic Acids; Address; Air; Bioinformatics; Biological Assay; COVID-19 detection; CRISPR/Cas technology; Cancer Etiology; Cellular Phone; Cervical Cancer Screening; Cessation of life; Chemicals; Clinical; Clinical Sensitivity; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Cytology; DNA; Data Reporting; Detection; Devices; Engineering; Ensure; Equipment; Eye; Feedback; Funding; Genotype; Hand; Health; Health Personnel; Human Papillomavirus; Human papilloma virus infection; Image; Infectious Diseases Research; Interdisciplinary Study; Laboratories; Lesion; Malignant neoplasm of cervix uteri; Methods; Microfluidic Microchips; Microfluidics; Molecular; Nucleic Acids; Oncologist; Optics; Paper; Performance; Polymerase Chain Reaction; Printing; Procedures; Process; Published Comment; RNA; Reaction; Reporting; Reproducibility; Resource-limited setting; Resources; Sampling; Sensitivity and Specificity; Services; Specificity; Speed; Swab; System; Technology; Temperature; Test Result; Testing; Time; Training; United States Food and Drug Administration; Virus; Visual; Waxes; Woman; World Health Organization; Zambia; anticancer research; base; burden of illness; chronic infection; clinical application; cost; design; detection sensitivity; detector; diagnostic platform; emerging pathogen; experience; global health; health training; high risk; industry partner; innovation; instrument; internal control; low and middle-income countries; multiplex detection; novel; point of care; point-of-care diagnostics; portability; rapid technique; smartphone Application

Abstract Cervical cancer is among the leading causes of cancer death in women worldwide, especially in low- and middle- income countries (LMICs). High-risk human papillomaviruses (HPV) are the main causative agents of cervical cancer and its precursor lesions; therefore, the World Health Organization (WHO) has recommended HPV DNA testing for cervical cancer screening in LMICs. Although polymerase chain reaction (PCR) methods have been widely used for HPV DNA detection, they are restricted to centralized clinical laboratories due to the need for labor-intensive procedures and expensive equipment. Here, we propose to develop a simple, rapid, highly sensitive, and specific CRISPR-on-paper diagnostic platform to simultaneously detect multiple high-risk HPV genotypes for cervical cancer screening at the point of care. This innovative diagnostic system is based on our recently developed all-in-one dual CRISPR-Cas12a (AIOD-CRISPR) assay, which combines the simplicity and high sensitivity of isothermal nucleic acid amplification with the high specificity of CRISPR detection. To develop a low-cost, multiplexed molecular detection technology, we will incorporate the AIOD-CRISPR assay into a paper-based microfluidics platform. To eliminate the need for complex electronic instruments, we will take advantage of an exothermic reaction to generate chemical heat for the CRISPR-on-paper system by using a disposable hand warmer, thus enabling instrument-free cervical cancer screening. The detection results can be read by the naked eye or reported by a programmed smartphone without the need for an expensive optical detector. We will rigorously evaluate and validate the clinical applications of our CRISPR-on-paper diagnostic system by testing clinical samples in collaboration with clinicians and healthcare workers in UConn Health and Zambia. If successful, the proposed project has an important impact on global health by providing a simple, affordable, and sensitive method for rapid screening of cervical cancer in resource-poor settings. As a platform technology, the proposed CRISPR-on-paper diagnostic system can be easily adapted to detect other emerging pathogens.

摘要 宫颈癌是全球女性癌症死亡的主要原因之一，特别是在中低收入人群中。 低收入国家（LMIC）。高危型人乳头瘤病毒（HPV）是宫颈癌的主要病原体， 癌症及其前驱病变;因此，世界卫生组织（WHO）建议HPV DNA 在中低收入国家进行宫颈癌筛查。虽然聚合酶链反应（PCR）方法已经被广泛应用于临床。 广泛用于HPV DNA检测，但由于需要 劳动密集型程序和昂贵的设备。在这里，我们建议开发一个简单，快速，高度 敏感、特异的CRISPR纸上诊断平台，可同时检测多种高危HPV 宫颈癌筛查的基因型在护理点。这种创新的诊断系统是基于我们的 最近开发的一体化双重CRISPR-Cas 12 a（AIOD-CRISPR）测定，其结合了简单性和 等温核酸扩增的高灵敏度与CRISPR检测的高特异性。发展 作为一种低成本、多路复用的分子检测技术，我们将把AIOD-CRISPR检测技术整合到 纸基微流体平台。为了消除对复杂电子仪器的需求，我们将采取 通过使用CRISPR-纸上系统产生化学热的放热反应的优点 一次性暖手器，从而实现无仪器宫颈癌筛查。检测结果可以 通过肉眼读取或通过编程的智能手机报告，而无需昂贵的光学传感器。 检测器我们将严格评估和验证我们的CRISPR纸上诊断的临床应用， 通过与康州大学健康中心的临床医生和医护人员合作测试临床样本， 赞比亚.如果成功，拟议的项目将对全球健康产生重要影响， 在资源匮乏的环境中，为快速筛查宫颈癌提供负担得起的敏感方法。为平台 技术，提出的CRISPR纸上诊断系统可以很容易地适应检测其他新兴的 病原体