A Precise and Portable Detection System for Infectious Pathogens Based on CRISPR/Cas Technology
10.19756/j.issn.0253-3820.231382
- VernacularTitle:基于CRISPR/Cas技术的传染性病原精准便携化检测系统
- Author:
Yi-Chen LIU
1
,
2
;
Ru-Jian ZHAO
;
Bai-Yang LYU
;
De-Feng SONG
;
Yi-Dan TANG
;
Yan-Fang JIANG
;
Bing-Ling LI
Author Information
1. 中国科学技术大学应用化学与工程学院,合肥 230026
2. 中国科学院长春应用化学研究所,电分析化学国家重点实验室,长春 130022
- Keywords:
Isothermal amplification;
CRISPR-Cas;
Fluorescence;
Portable detection;
Infectious pathogens;
Grayscale reader
- From:
Chinese Journal of Analytical Chemistry
2024;52(2):187-197
- CountryChina
- Language:Chinese
-
Abstract:
Nucleic acid-based molecular diagnostic methods are considered the gold standard for detecting infectious pathogens.However,when applied to portable or on-site rapid diagnostics,they still face various limitations and challenges,such as poor specificity,cumbersome operation,and portability difficulties.The CRISPR(Clustered regularly interspaced short palindromic repeats)/CRISPR-associated protein(Cas)-fluorescence detection method holds the potential to significantly enhance the specificity and signal-to-noise ratio of nucleic acid detection.In this study,we developed a portable grayscale reader detection system based on loop-mediated isothermal amplification(LAMP)-CRISPR/Cas.On one hand,in the presence of CRISPR RNA(crRNA),the CRISPR/Cas12a system was employed to achieve precise fluorescent detection of self-designed LAMP amplification reactions for influenza A and influenza B viruses.This further validated the high selectivity and versatility of the CRISPR/Cas system.On the other hand,the accompanying independently developed portable grayscale reader allowed for low-cost collection of fluorescence signals and high-reliability visual interpretation.At the end of the detection process,it directly provided positive or negative results.Practical sample analyses using this detection system have verified its reliability and utility,demonstrating that this system can achieve highly sensitive and highly specific portable analysis of influenza viruses.
