Bang D D, Wedderkopp A, Pedersen K, Madsen M
Department of Poultry, Fish, and Fur Animals, Danish Veterinary Institute, Hangøvej 2, DK-8200 Aarhus N, Denmark.
Mol Cell Probes. 2002 Oct;16(5):359-69. doi: 10.1006/mcpr.2002.0434.
Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited sensitivity due to the use of selective media, the low number of bacteria in the samples and possibly also due to the presence of non-culturable or sub-lethally injured stages of the bacteria. The present paper describes a rapid PCR assay using nested primers of the 16S rRNA or the hippuricase (hip O) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples. The sensitivity of the nested PCR was determined to be 0.01 pg/PCR, corresponding to 2-3 colony forming units (cfu) per ml. The nested PCR assays were applied to detect C. jejuni and C. coli in 269 environmental samples collected from ten broiler farms. The sensitivity, specificity and the usefulness of the PCR assay for detection of C. jejuni and C. coli in environmental samples are presented and discussed.
家禽养殖场弯曲杆菌感染源的识别通常存在问题,因为缺乏可靠的方法来检测环境样本中的弯曲杆菌。由于使用选择性培养基、样本中细菌数量少以及可能还由于存在细菌的不可培养或亚致死损伤阶段,通过传统培养方法检测环境样本中的弯曲杆菌很困难且灵敏度有限。本文描述了一种使用16S rRNA或马尿酸酶(hip O)基因的巢式引物来检测环境样本中空肠弯曲杆菌和结肠弯曲杆菌的快速PCR检测方法。巢式PCR的灵敏度确定为0.01 pg/PCR,相当于每毫升2-3个菌落形成单位(cfu)。将巢式PCR检测方法应用于检测从十个肉鸡养殖场采集的269份环境样本中的空肠弯曲杆菌和结肠弯曲杆菌。文中展示并讨论了该PCR检测方法在检测环境样本中空肠弯曲杆菌和结肠弯曲杆菌时的灵敏度、特异性及实用性。