title: Comparative quantification of Campylobacter jejuni from environmental samples using traditional and molecular biological techniques.
abstract: Campylobacter jejuni is one of the most common causes of gastroenteritis in the world. Given the potential risks to human, animal, and environmental health, the development and optimization of methods to quantify this important pathogen in environmental samples is essential. Two of the most commonly used methods for quantifying C. jejuni are selective plate counting and quantitative real-time PCR (qPCR). Unfortunately, little comparative research has been performed to evaluate the accuracy of these methods for quantification of C. jejuni in aqueous and solid matricies. In this study, the limit of detection and the level of resolution obtained using these 2 methods was evaluated for C. jejuni and compared with that of the common indicator organism Escherichia coli. The use of selective plate count media for quantification of C. jejuni resulted in a 0.7-1.2 log underestimation of cell concentrations, compared with qPCR in both water and column leachate samples, whereas E. coli concentrations were found to be similar with either technique. For C. jejuni, only the qPCR assay accurately measured 2-fold changes in cell concentrations in water samples, whereas concentrations of E. coli were accurately measured regardless of method. Based on these data, qPCR assays were found to be more accurate than selective plate counts for quantification of C. jejuni from environmental samples.
authors.affiliation: USDA-ARS, Animal Waste Management Research Unit (AWMRU), Bowling Green, KY 42104, USA. michael.rothrock@ars.usda.gov
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Rothrock
authors.forename: Kimberly L
authors.initials: KL
authors.lastname: Cook
authors.forename: Carl H
authors.initials: CH
authors.lastname: Bolster
mesh_terms: D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial / Q000379:methods*; D004783:Environmental Microbiology; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D008967:Molecular Biology / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity