Adapted method for rapid detection and quantification of pathogen Campylobacter jejuni from environmental water samples.
FEMS Microbiol Ecol
; 99(7)2023 06 16.
Article
in En
| MEDLINE
| ID: mdl-37245057
ABSTRACT
Building on a previously developed workflow for rapid and sensitive pathogen detection by qPCR, this work has established a sample treatment strategy that produces consistent quantification efficiencies (QEs) for Campylobacter jejuni against a complex and highly variable sample matrix from a suburban river. The individual treatments most effective at minimizing the inhibitory effects of the sample matrix were pH buffering with HEPES (50 mM, pH 5.7) and addition of the surfactant Tween 20 (2% v/v). Unexpectedly, sample acidification (pH 4-5) resulting from the use of aged Tween 20 that had undergone partial hydrolysis, appeared to play a key role in enhancing QE. This effect could be replicated by direct pH adjustment with dilute hydrochloric acid and may be linked to the solubilization and removal of inhibitory particles at an acidic pH. While the effectiveness of each individual treatment method varied, a combined treatment of either HEPES buffer + Tween 20, or direct pH adjustment + Tween 20, consistently produced QEs of 60%-70% and up to 100%, respectively, over a sampling period of one year. The consistency and scalability of this workflow make it a suitable alternative to culture-based ISO methods for detecting Campylobacter spp.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Campylobacter jejuni
/
Campylobacter coli
Type of study:
Diagnostic_studies
Language:
En
Journal:
FEMS Microbiol Ecol
Year:
2023
Document type:
Article
Affiliation country:
Australia