RPA-CRISPR-Cas 介导的双侧向层析检测用于 HPV16 和 HPV18 的即时检测。
RPA-CRISPR-Cas-Mediated Dual Lateral Flow Assay for the Point-of-Care Testing of HPV16 and HPV18.
发表日期:2024 Oct 15
作者:
Kaini Zhang, Qingmei Li, Kun Wang, Qiaoli Zhang, Chengkun Ma, Guiwen Yang, Yanxia Xie, Michael G Mauk, Shanji Fu, Lei Chen
来源:
BIOCONJUGATE CHEMISTRY
摘要:
近年来,由人乳头瘤病毒(HPV)感染引起的宫颈癌发病率有所上升。超过一半的宫颈癌病例是由 HPV16 和 HPV18 感染引起的,因此 HPV16 和 HPV18 检测对于预防宫颈癌至关重要。 HPV检测主要在医院进行,但受时间和专业医疗设施的限制。另一方面,如果能够实现足够的灵敏度和特异性,由于隐私性和时间和地点方面的灵活性,使用简单诊断的家庭自我测试将是一种有吸引力的替代方案。在这项工作中,描述了一种基于 RPA-CRISPR-Cas12a/13a(称为 RC-LFA)的用于 HPV 检测的双侧流分析。利用Cas12a和Cas13a的切割特异性,设计了CRISPR-Cas12a/Cas13a系统来检测HPV16和HPV18。具有两条测试线的侧流试纸条专为适合 CRISPR-Cas12a/Cas13 系统而设计。 RC-LFA实现了HPV16和HPV18的快速同步检测,具有高特异性和灵敏度(10拷贝/μL),从核酸提取到免仪器读数仅需约40分钟。 RC-LFA 用户友好且无需仪器,使其成为一种很有前途的家庭 HPV 自检方法。
The incidence of cervical cancer caused by human papillomavirus (HPV) infection has increased in recent years. More than half of all cervical cancer cases are due to HPV16 and HPV18 infection, so HPV16 and HPV18 testing is essential to prevent cervical cancer. HPV testing is mainly carried out in hospitals, but it is subject to time and specialized medical facilities. On the other hand, home self-testing using simple diagnostics would present an attractive alternative due to privacy and flexibility with regard to time and place, provided sufficient sensitivity and specificity can be achieved. In this work, a dual lateral flow assay based on RPA-CRISPR-Cas12a/13a (named RC-LFA) for HPV detection was described. Taking advantage of the cleavage specificity of Cas12a and Cas13a, a CRISPR-Cas12a/Cas13a system was designed to detect HPV16 and HPV18. The lateral flow strip with two test lines was designed to suit the CRISPR-Cas12a/Cas13 system. RC-LFA achieves rapid and simultaneous detection of HPV16 and HPV18 with high specificity and sensitivity (10 copies/μL) in about 40 min from the extraction of nucleic acid to an instrument-free readout. RC-LFA is user-friendly and instrument-free, making it a promising method for HPV self-tests at home.