Multispectral pathogens detection in food using multiplex hyperbranched saltatory rolling circle amplification.
Talanta
; 279: 126618, 2024 Nov 01.
Article
in En
| MEDLINE
| ID: mdl-39116729
ABSTRACT
Foodborne illnesses caused by Salmonella and Staphylococcus aureus are a significant public health concern, leading to societal and economic repercussions. It is important to develop a simple and straightforward bacteria detection and identification method. A triple-probe multiplex rolling circle amplification technique has been developed to simultaneously detect Salmonella Typhimurium and S. aureus. This method utilizes fluorophore-labeled long padlock probes targeting S. Typhimurium invA and S. aureus glnA specific genes, along with a pH-based detection approach for direct visual identification. The multiplex hyperbranched saltatory rolling circle amplification assay at 30 °C has showed promising results with synthetic targets within 30 min and real bacteria within 2 h after establishing the detection settings. The assay is specific for S. aureus and S. Typhimurium, with a limit of detection of 39 µM for fluorescence and 78 µM for colorimetric. In the simulative test of this method for the detection of S. Typhimurium and S. aureus in milk, the limit of detection for the fluorescence signal after 2 h of amplification was 10 CFU/mL and 5 CFU/mL, respectively. The detection method was evaluated to be stable enough to detect pathogen for 3.29 months. Consequently, this triple-probe-multiplex rolling circle amplification method displays notable specificity, sensitivity, as well as ease of interpretation when testing food samples for harmful pathogens.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Salmonella typhimurium
/
Staphylococcus aureus
/
Nucleic Acid Amplification Techniques
/
Food Microbiology
Limits:
Animals
Language:
En
Journal:
Talanta
Year:
2024
Document type:
Article
Country of publication:
Netherlands