Cyclic Multiple Primer Generation Rolling Circle Amplification Assisted Capillary Electrophoresis for Simultaneous and Ultrasensitive Detection of Multiple Pathogenic Bacteria

Efficient, accurate, and economical detection of pathogenic bacteria is crucial in ensuring food safety and preventing foodborne illnesses. How to fulfill the highly sensitive and simultaneous detection of multiple trace pathogenic bacteria is a big challenge. In this work, capillary electrophoresis...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2024-01, Vol.96 (4), p.1781-1788
Main Authors: Chu, Zhaohui, Chen, Jingyi, Zhang, Jingzi, Xie, Qihui, Zhang, Fan, Wang, Qingjiang
Format: Article
Language:English
Subjects:
Amplification
Bacteria
Capillary electrophoresis
Deoxyribonucleic acid
DNA
Drinking water
Electrophoresis
Food safety
Foodborne diseases
Salmonella enterica enterica
Salmonella Typhi
Shigella dysenteriae
Vibrio parahaemolyticus
Water analysis
Water sampling
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Summary:Efficient, accurate, and economical detection of pathogenic bacteria is crucial in ensuring food safety and preventing foodborne illnesses. How to fulfill the highly sensitive and simultaneous detection of multiple trace pathogenic bacteria is a big challenge. In this work, capillary electrophoresis coupled with a cyclic multiple primer generation rolling circle amplification (cyclic MPG-RCA) was studied for highly sensitive and simultaneous detection of three kinds of pathogenic bacteria. The cyclic MPG-RCA was based on a carefully designed clover-shaped DNA probe, in which three “leaves” corresponded to three types of aimed pathogenic bacteria: Shigella dysenteriae (S. dysenteriae), Salmonella enterica subsp. enterica serovar Typhi (S. Typhi), and Vibrio parahaemolyticus (V. parahaemolyticus). Under the optimal experimental conditions, the limits of detection (S/N = 3) of this method for bacterial target DNA were 11.4 amol·L–1 (S. dysenteriae), 4.88 amol·L–1 (S. Typhi), and 14.9 amol·L–1 (V. parahaemolyticus), and the conversion concentrations for the target bacteria were 10 colony-forming units (CFU)·mL–1 (S. dysenteriae), 3 CFU·mL–1 (S. Typhi), and 12 CFU·mL–1 (V. parahaemolyticus). This method had been applied to the detection of tap water samples with good results, which proved that it could be used as an effective tool for trace pathogenic bacteria monitoring in foods, environments, and medicines.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.3c05117