通过一种新型多重 PCR 检测方法鉴定五种人类和哺乳动物相关的弯曲菌属物种。
Identification of five human and mammal associated Arcobacter species by a novel multiplex-PCR assay.
机构信息
Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
出版信息
J Microbiol Methods. 2010 Mar;80(3):281-6. doi: 10.1016/j.mimet.2010.01.009. Epub 2010 Jan 21.
A multiplex-PCR assay with seven primers was developed for the identification of the five human and mammal related species of the emerging foodborne pathogen Arcobacter. The assay was validated using 58 reference and 358 collection strains isolated from humans and mammals. The selected primers on the 23 S RNA gene amplify a 2061 bp fragment from A. butzleri, a 1590 bp fragment from A. thereuis, a 1125 bp fragment from A. cibarius and an A. skirrowii specific fragment of 198 bp. For A. cryaerophilus, a primer set on the gyrA gene amplified a specific fragment of 395 bp. No PCR product was generated for closely related bacteria including Campylobacter and Helicobacter species. Furthermore, examination of the 23 S RNA gene of A. cryaerophilus revealed, besides large heterogeneity, the presence of intervening sequences ranging from 87 to 196 bp.
建立了一种使用七对引物的多重 PCR 检测方法，用于鉴定新兴食源性致病菌弯曲弧菌属中与人类和哺乳动物相关的五个种。该方法使用了 58 株参考菌株和 358 株从人类和哺乳动物中分离得到的菌株进行了验证。所选的 23S rRNA 基因上的引物可从 A. butzleri 中扩增出 2061bp 的片段，从 A. thereuis 中扩增出 1590bp 的片段，从 A. cibarius 中扩增出 1125bp 的片段，从 A. skirrowii 中扩增出 198bp 的特异性片段。对于 A. cryaerophilus，gyrA 基因上的引物组可扩增出 395bp 的特异性片段。与该方法密切相关的细菌，包括弯曲菌属和幽门螺杆菌属，均未产生 PCR 产物。此外，对 A. cryaerophilus 的 23S rRNA 基因的研究表明，除了存在较大的异质性外，还存在 87-196bp 不等的插入序列。
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