RPA-CRISPR-CAS介导的双侧向流量测定HPV16和HPV18的护理点测试

RPA-CRISPR-Cas-Mediated Dual Lateral Flow Assay for the Point-of-Care Testing of HPV16 and HPV18

BIOCONJUGATE CHEMISTRY

影响因子:3.90000

分区:化学2区 / 生化研究方法2区有机化学2区生化与分子生物学3区化学：综合3区

发表日期:2024 Nov 20

作者: Kaini Zhang, Qingmei Li, Kun Wang, Qiaoli Zhang, Chengkun Ma, Guiwen Yang, Yanxia Xie, Michael G Mauk, Shanji Fu, Lei Chen

摘要

近年来，由人乳头瘤病毒（HPV）感染引起的宫颈癌的发生率增加。在所有宫颈癌病例中，超过一半是由于HPV16和HPV18感染引起的，因此HPV16和HPV18测试对于预防宫颈癌至关重要。 HPV测试主要在医院进行，但要遵守时间和专门的医疗设施。另一方面，使用简单诊断的家庭进行自我测试将是一个有吸引力的替代方法，因为在时间和地点方面的灵活性，只要可以实现足够的敏感性和特异性。在这项工作中，描述了基于RPA-CRISPR-CAS12A/13A（称为RC-LFA）HPV检测的双侧向流量测定。利用CAS12A和CAS13A的裂解特异性，CRISPR-CAS12A/CAS13A系统旨在检测HPV16和HPV18。具有两条测试线的侧向流带设计适合CRISPR-CAS12A/CAS13系统。 RC-LFA可在大约40分钟内从核酸提取到无仪器读数的大约40分钟内，在大约40分钟内以高特异性和灵敏度（10拷贝/μL）快速和同时检测HPV16和HPV18。 RC-LFA是用户友好且不含仪器的，使其成为HPV自我测试的有前途的方法。

Abstract

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.