Electrochemical biosensor for detecting pathogenic bacteria based on a hybridization chain reaction and CRISPR-Cas12a

In this study, Lba Cas12a (Cpf1) as one of the CRISPR systems from Lachnospiraceae bacterium was coupled with a hybridization chain reaction (HCR) to develop an electrochemical biosensor for detecting the pathogenic bacterium, Salmonella typhimurium . Autonomous cross-opening of functional DNA hairp...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2022-01, Vol.414 (2), p.1073-1080
Main Authors: Liu, Xiu, Bu, Shengjun, Feng, Jingqi, Wei, Hongguo, Wang, Ze, Li, Xue, Zhou, Hongyu, He, Xiuxia, Wan, Jiayu
Format: Article
Language:English
Subjects:
Analysis
Analytical Chemistry
Aptamers
Bacteria
Biochemistry
Biosensing Techniques - methods
Biosensors
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
CRISPR
CRISPR-Cas Systems
Deoxyribonucleic acid
Detection limits
DNA
Electrochemical apparatus
Electrochemical Techniques - methods
Electrochemistry
Electron transfer
Electrophoresis, Polyacrylamide Gel
Food Science
Genetic aspects
Hybridization
Identification and classification
Laboratory Medicine
Methods
Monitoring/Environmental Analysis
Polymers
Research Paper
Salmonella
Salmonella typhimurium
Salmonella typhimurium - isolation & purification
Salmonella typhimurium - pathogenicity
Single-stranded DNA
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Summary:In this study, Lba Cas12a (Cpf1) as one of the CRISPR systems from Lachnospiraceae bacterium was coupled with a hybridization chain reaction (HCR) to develop an electrochemical biosensor for detecting the pathogenic bacterium, Salmonella typhimurium . Autonomous cross-opening of functional DNA hairpin structures of HCR yielded polymer double-stranded DNA wires consisting of numerous single-stranded DNAs, which initiated the trans-cleavage activity of CRISPR-Cas12a to indiscriminately cleave random single-stranded DNA labeling electrochemical tags on the surface of the electrode. It led to a variation in the electron transfer of electrochemical tags. The polymer double-stranded DNA of HCR was immobilized on dynabeads (DBs) via the S. typhimurium aptamer and released from DBs. The established method could selectively and sensitively quantify S. typhimurium in samples with detection limits of 20 CFU/mL. Our study provides a novel insight for exploring universal analytical methods for pathogenic bacteria based on CRISPR-Cas12a coupled with HCR.
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-021-03733-6