Unitat de Microbiologia, Departament de Ciències Mediques Bàsiques, Facultat de Medicina i Ciències de la Salut, IISPV, Universitat Rovira i Virgili, Reus, Spain.
BMC Microbiol. 2012 Dec 18;12:292. doi: 10.1186/1471-2180-12-292.
Arcobacter spp. (family Campylobacteraceae) are ubiquitous zoonotic bacteria that are being increasingly recognised as a threat to human health. A previously published 16S rRNA-RFLP Arcobacter spp. identification method produced specific RFLP patterns for the six species described at that time, using a single endonuclease (MseI). The number of characterised Arcobacter species has since risen to 17. The aim of the present study was to update the 16S rRNA-RFLP identification method to include all currently characterised species of Arcobacter.
Digestion of the 16S rRNA gene with the endonuclease MseI produced clear, distinctive patterns for 10 of the 17 species, while the remaining species shared a common or very similar RFLP pattern. Subsequent digestion of the 16S rRNA gene from these species with the endonucleases MnlI and/or BfaI generated species-specific RFLP patterns.
16S rRNA-RFLP analysis identified 17 Arcobacter spp. using either polyacrylamide or agarose gel electrophoresis. Microheterogeneities within the 16S rRNA gene, which interfered with the RFLP identification, were also documented for the first time in this genus, particularly in strains of Arcobacter cryaerophilus isolated from animal faeces and aborted foetuses.
弯曲杆菌属(弯曲菌科)是普遍存在的人畜共患病细菌,它们对人类健康的威胁正日益受到关注。先前发表的 16S rRNA-RFLP 弯曲杆菌属鉴定方法使用单一内切酶(MseI)为当时描述的六种物种产生了特定的 RFLP 图谱。此后,被鉴定的弯曲杆菌属物种数量已增加到 17 种。本研究的目的是更新 16S rRNA-RFLP 鉴定方法,以包括所有目前已鉴定的弯曲杆菌属物种。
用内切酶 MseI 消化 16S rRNA 基因为 10 种 17 种物种产生了清晰、独特的图谱,而其余物种具有共同或非常相似的 RFLP 图谱。随后,用内切酶 MnlI 和/或 BfaI 消化这些物种的 16S rRNA 基因,产生了物种特异性的 RFLP 图谱。
16S rRNA-RFLP 分析使用聚丙烯酰胺或琼脂糖凝胶电泳鉴定了 17 种弯曲杆菌属。本研究还首次在该属中记录了 16S rRNA 基因内的微异质性,这会干扰 RFLP 鉴定,尤其是在从动物粪便和流产胎儿中分离的嗜冷弯曲杆菌菌株中。