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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的宫颈癌是全世界女性癌症死亡的主要原因之一，特别是在低收入和中等收入女性中收入国家（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期病变；因此，世界卫生组织（WHO）推荐HPV DNA 中低收入国家宫颈癌筛查测试。尽管聚合酶链式反应 (PCR) 方法已广泛用于 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 用于护理时宫颈癌筛查的基因型。这种创新的诊断系统基于我们的最近开发的一体化双 CRISPR-Cas12a (AIOD-CRISPR) 检测方法，结合了简单性和等温核酸扩增的高灵敏度和CRISPR检测的高特异性。发展作为一种低成本、多重分子检测技术，我们将把 AIOD-CRISPR 检测结合到基于纸的微流控平台。为了消除对复杂电子仪器的需求，我们将采取利用放热反应为 CRISPR-on-paper 系统产生化学热一次性暖手器，从而实现无仪器宫颈癌筛查。检测结果可通过肉眼读取或通过编程的智能手机报告，无需昂贵的光学设备探测器。我们将严格评估和验证我们的 CRISPR 纸上诊断的临床应用系统通过与康涅狄格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，拟议的项目将通过提供简单的、在资源匮乏的环境中快速筛查宫颈癌的经济且灵敏的方法。作为一个平台技术，所提出的 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。