Environmental monitoring of waterborne Campylobacter: evaluation of the Australian Standard and a hybrid extraction-free MPN-PCR method.

• Main Authors: Rebekah eHenry, Christelle eSchang, gayani echandrasena, Ana eDeletic, Mark eEdmunds, dusan ejovanovic, peter ekolotelo, richard ewilliamson, Jonathan eSchmidt, David eMcCarthy
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2015-02-01
• Series: Frontiers in Microbiology
• Subjects: Campylobacter; PCR; environmental interactions; estuary; culture.; Inter-laboratory
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fmicb.2015.00074/full

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.