Henry Rebekah, Schang Christelle, Chandrasena Gayani I, Deletic Ana, Edmunds Mark, Jovanovic Dusan, Kolotelo Peter, Schmidt Jonathan, Williamson Richard, McCarthy David
Environmental and Public Health Laboratory, Department of Civil Engineering, Monash University Clayton, VIC, Australia.
ALS Environmental Scoresby, VIC, Australia.
Front Microbiol. 2015 Feb 9;6:74. doi: 10.3389/fmicb.2015.00074. eCollection 2015.
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.
弯曲杆菌是全球腹泻疾病的主要病原体。本研究评估了一种新型的培养 - PCR杂交（MPN - PCR）检测方法，用于快速计数来自河口和废水系统中的弯曲杆菌属。为首先评估当前基于培养的澳大利亚标准，对69个分样水样进行了一项实验室间研究。然后通过分析在两年期间收集的147个河口样本，对提议的最大可能数（MPN）- PCR方法进行了评估。还整理了14种不同的生物学、水文和气候参数的数据，以确定病原体与环境的关系，并评估方法特异性偏差的可能性。结果表明，MPN - PCR的实验室内性能优于AS/NZS（σ = 0.7912，P < 0.001；κ = 0.701，P < 0.001），总体诊断准确率约为94%。此外，对MPN - PCR和AS/NZS的分析均表明，在评估环境参数对弯曲杆菌属数量的影响时，存在引入方法特异性偏差的可能性。
