title: Environmental monitoring of waterborne Campylobacter: evaluation of the Australian standard and a hybrid extraction-free MPN-PCR method.
abstract: Campylobacter is the leading agent of diarrheal disease worldwide. This study evaluates a novel culture-PCR hybrid (MPN-PCR) assay for the rapid enumeration of Campylobacter spp. from estuarine and wastewater systems. To first evaluate the current, culture-based, Australian standard, an inter-laboratory study was conducted on 69 subsampled water samples. The proposed Most-Probable Number (MPN)-PCR method was then evaluated, by analysing 147 estuarine samples collected over a 2 year period. Data for 14 different biological, hydrological and climatic parameters were also collated to identify pathogen-environment relationships and assess the potential for method specific bias. The results demonstrated that the intra-laboratory performance of the MPN-PCR was superior to that of AS/NZS (σ = 0.7912, P < 0.001; κ = 0.701, P < 0.001) with an overall diagnostic accuracy of ~94%. Furthermore, the analysis of both MPN-PCR and AS/NZS identified the potential for the introduction of method specific bias during assessment of the effects of environmental parameters on Campylobacter spp. numbers.
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Rebekah
authors.initials: R
authors.lastname: Henry
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Christelle
authors.initials: C
authors.lastname: Schang
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Gayani I
authors.initials: GI
authors.lastname: Chandrasena
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Ana
authors.initials: A
authors.lastname: Deletic
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Mark
authors.initials: M
authors.lastname: Edmunds
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Dusan
authors.initials: D
authors.lastname: Jovanovic
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: Kolotelo
authors.affiliation: ALS Environmental Scoresby, VIC, Australia.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Schmidt
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: Richard
authors.initials: R
authors.lastname: Williamson
authors.affiliation: Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
authors.forename: David
authors.initials: D
authors.lastname: McCarthy
keywords: Campylobacter; PCR; culture; environmental interactions; estuary; inter-laboratory