Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
J Food Prot. 2010 Jun;73(6):1057-63. doi: 10.4315/0362-028x-73.6.1057.
We tested the use of multiplex real-time PCR for detection and quantification of Campylobacter jejuni and Campylobacter coli on broiler carcass neck skin samples collected during 2008 from slaughterhouses in Switzerland. Results from an established TaqMan assay based on two different targets (hipO and ceuE for C. jejuni and C. coli, respectively) were corroborated with data from a newly developed assay based on a single-nucleotide polymorphism in the fusA gene, which allows differentiation between C. jejuni and C. coli. Both multiplex real-time PCRs were applied simultaneously for direct detection, differentiation, and quantification of Campylobacter from 351 neck skin samples and compared with culture methods. There was good correlation in detection and enumeration between real-time PCR results and quantitative culture, with real-time PCR being more sensitive. Overall, 251 (71.5%) of the samples were PCR positive for Campylobacter, with 211 (60.1%) in the hipO-ceuE assays, 244 (69.5%) in the fusA assay, and 204 (58.1%) of them being positive in both PCR assays. Thus, the fusA assay was similarly sensitive to the enrichment culture (72.4% positive); however, it is faster and allows for quantification. In addition, real-time PCR allowed for species differentiation; roughly 60% of positive samples contained C. jejuni, less than 10% C. coli, and more than 30% contained both species. Real-time PCR proved to be a suitable method for direct detection, quantification, and differentiation of Campylobacter from carcasses, and could permit time-efficient surveillance of these zoonotic agents.
我们测试了多重实时 PCR 检测和定量分析 2008 年瑞士屠宰场采集的肉鸡颈皮样本中空肠弯曲菌和结肠弯曲菌的方法。该方法基于两个不同靶标（hipO 和 ceuE 分别用于检测空肠弯曲菌和结肠弯曲菌）的已建立 TaqMan 检测结果，与基于 fusA 基因单核苷酸多态性建立的新检测方法的数据相符，后者可区分空肠弯曲菌和结肠弯曲菌。这两种多重实时 PCR 同时用于直接检测、区分和定量分析 351 个颈皮样本中的弯曲菌，并与培养方法进行了比较。实时 PCR 结果与定量培养在检测和计数方面相关性良好，实时 PCR 更敏感。总体而言，251 份（71.5%）样本的弯曲菌 PCR 检测呈阳性，其中 hipO-ceuE 检测 211 份（60.1%），fusA 检测 244 份（69.5%），两种 PCR 检测均为阳性的有 204 份（58.1%）。因此，fusA 检测与富集培养的灵敏度相似（阳性率为 72.4%）；然而，它更快并且可以定量。此外，实时 PCR 允许进行种属区分；大约 60%的阳性样本含有空肠弯曲菌，不到 10%的样本含有结肠弯曲菌，超过 30%的样本同时含有这两种菌。实时 PCR 被证明是一种直接从胴体中检测、定量和区分弯曲菌的合适方法，可用于有效监测这些人畜共患病原体。
