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

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Bioconjugate Chemistry Bioconjugate Pub Date : 2024-10-15 DOI:10.1021/acs.bioconjchem.4c0037510.1021/acs.bioconjchem.4c00375

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

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引用次数: 0

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.

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RPA-CRISPR-Cas 介导的双侧流检测法用于 HPV16 和 HPV18 的床旁检测

近年来，由人类乳头瘤病毒（HPV）感染引起的宫颈癌发病率有所上升。在所有宫颈癌病例中，有一半以上是由 HPV16 和 HPV18 感染所致，因此，HPV16 和 HPV18 检测对预防宫颈癌至关重要。HPV 检测主要在医院进行，但受到时间和专业医疗设施的限制。另一方面，如果能达到足够的灵敏度和特异性，利用简单的诊断方法进行家庭自我检测将是一种有吸引力的替代方法，因为它具有隐私性和时间地点的灵活性。在这项工作中，描述了一种基于 RPA-CRISPR-Cas12a/13a 的双侧流检测法（命名为 RC-LFA），用于检测 HPV。利用 Cas12a 和 Cas13a 的裂解特异性，设计了一个 CRISPR-Cas12a/Cas13a 系统来检测 HPV16 和 HPV18。为适应 CRISPR-Cas12a/Cas13 系统，设计了带有两条检测线的侧流试纸。RC-LFA 可同时快速检测 HPV16 和 HPV18，特异性和灵敏度高（10 拷贝/μL），从提取核酸到无仪器读出约需 40 分钟。RC-LFA 操作简便，无需仪器，是一种很有前景的家庭 HPV 自我检测方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioconjugate Chemistry Bioconjugate 生物-化学综合

CiteScore

9.00

自引率

2.10%

发文量

236

审稿时长

1.4 months

期刊介绍： Bioconjugate Chemistry invites original contributions on all research at the interface between man-made and biological materials. The mission of the journal is to communicate to advances in fields including therapeutic delivery, imaging, bionanotechnology, and synthetic biology. Bioconjugate Chemistry is intended to provide a forum for presentation of research relevant to all aspects of bioconjugates, including the preparation, properties and applications of biomolecular conjugates.