Poly(vinylpyrrolidone)-Enhanced CRISPR-Cas System for Robust Nucleic Acid Diagnostics

Clustered regularly interspaced short palindromic repeats (CRISPR) technology has opened a new path for molecular diagnostics based on RNA programmed trans-cleavage activity. However, their accessibility for highly sensitive clinical diagnostics remains insufficient. In this study, we systematically...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2024-10, Vol.96 (39), p.15797-15807
Main Authors: Liu, Pengfei, Zeng, Jiayu, Jiang, Chengchuan, Du, Jinlian, Jiang, Ling, Li, Sheng, Zeng, Fanxu, Xiong, Erhu
Format: Article
Language:English
Subjects:
CRISPR
Human papillomavirus
Medical research
Nucleic acids
Polymerase chain reaction
Polyvinylpyrrolidone
Surfactants
Vinylpyrrolidone
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Summary:Clustered regularly interspaced short palindromic repeats (CRISPR) technology has opened a new path for molecular diagnostics based on RNA programmed trans-cleavage activity. However, their accessibility for highly sensitive clinical diagnostics remains insufficient. In this study, we systematically investigated the impact of various surfactants on the CRISPR-Cas12a system and found that poly­(vinylpyrrolidone) (PVP), a nonionic surfactant, showed the highest enhancement effect among these tested surfactants. Additionally, the enhancement effects of PVP are compatible and versatile to CRISPR-Cas12b and Cas13a systems, improving the sensitivity of these CRISPR-Cas systems toward synthetic targets by 1–2 orders of magnitude. By integrating the PVP-enhanced CRISPR system with isothermal nucleic acid amplification, both the two- and one-step assays exhibited comparable sensitivity and specificity to gold-standard quantitative polymerase chain reaction (qPCR) in the assay of clinical human papillomavirus (HPV) samples, thereby holding significant promise for advancing clinical diagnostics and biomedical research.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.4c04077