O'Sullivan N A, Fallon R, Carroll C, Smith T, Maher M
DNA Diagnostics, National Diagnostics Centre, Galway, Ireland.
Mol Cell Probes. 2000 Feb;14(1):7-16. doi: 10.1006/mcpr.1999.0274.
Campylobacter enteritis in humans has been linked to consumption of poultry meat. Surveys show that 30-100% of poultry harbour Campylobacter as normal flora of the digestive tract which indicates a need to identify prevalent organism types in flocks and trace their epidemiology. In this study we describe a Campylobacter genus specific polymerase chain reaction (PCR) assay, amplifying the 16 S-23 S rRNA intergenic spacer region with an internal Campylobacter genus specific DNA probe and species specific probes for Campylobacter jejuni and Campylobacter coli designed for confirmation of the amplified PCR products by Southern blot and colorimetric reverse hybridization assays. The specificity of this assay was established by testing a range of food pathogens. Broiler chicken samples were tested following presumptive positive identification by the Malthus System V analyser (Malthus Instruments, UK). The combined PCR and colorimetric reverse hybridization assay is easy to perform and faster than conventional methods for confirmation and identification of Campylobacter species.
人类的弯曲杆菌肠炎与食用禽肉有关。调查显示，30%至100%的家禽携带弯曲杆菌作为消化道的正常菌群，这表明有必要识别鸡群中常见的菌株类型并追踪其流行病学情况。在本研究中，我们描述了一种弯曲杆菌属特异性聚合酶链反应（PCR）检测方法，该方法使用内部弯曲杆菌属特异性DNA探针扩增16 S - 23 S rRNA基因间隔区，并设计了空肠弯曲杆菌和结肠弯曲杆菌的种特异性探针，用于通过Southern印迹和比色反向杂交检测来确认扩增的PCR产物。通过检测一系列食品病原体确定了该检测方法的特异性。在通过马尔萨斯系统V分析仪（英国马尔萨斯仪器公司）初步阳性鉴定后，对肉鸡样本进行了检测。联合PCR和比色反向杂交检测方法易于操作，比传统方法更快地用于弯曲杆菌菌种的确认和鉴定。
