Animal Research Facility, Heinrich-Heine-University Duesseldorf, Duesseldorf, Germany.
J Microbiol Methods. 2012 Sep;90(3):342-9. doi: 10.1016/j.mimet.2012.06.013. Epub 2012 Jul 4.
The internal transcribed spacer (ITS) regions of members of Pasteurellaceae isolated from rodents, including the [Pasteurella] pneumotropica biotypes Jawetz and Heyl, [Actinobacillus] muris, "Hemophilus influenzaemurium" and Bisgaard taxon 17 were studied and their feasibility to discriminate these species was analyzed. The reference strains of all species analyzed showed unique species-specific ITS patterns which were further present in 49 clinical isolates of [P.] pneumotropica biotypes Jawetz and Heyl and [A.] muris allowing their identification by comparison to the reference strains pattern. Sequence analysis of the amplified fragments revealed in all species, with exception of "H. influenzaemurium", a larger ITS(ile+ala) which contained the genes for tRNA(Ile(GAU)) and tRNA(Ala(UGC)) and a smaller ITS(glu) with the tRNA(Glu(UUC)) gene. "H. influenzaemurium" revealed two each of the larger and respectively the smaller ITS fragments. Both the length and the sequence of each ITS type were highly conserved within the [P.] pneumotropica biotypes Jawetz and Heyl and [A.] muris strains tested. On the contrary, ITS sequences revealed significant interspecies variations with identity levels ranging from 61.2 to 89.5% for ITS(ile+ala) and 56.5 to 86.8% for ITS(glu). Sequences regions with significant interspecies variation but highly conserved within the species were identified and might be used to design probes for the identification of rodent Pasteurellaceae to the species level.
从啮齿动物中分离出的巴斯德氏菌科成员（包括巴氏杆菌气肿亚种 Jawetz 和 Heyl、鼠型嗜血杆菌、“流感嗜血杆菌 murium”和 Bisgaard 分类群 17）的内转录间隔区（ITS）区域进行了研究，并分析了其区分这些物种的可行性。分析的所有参考菌株均显示出独特的种特异性 ITS 模式，这些模式进一步存在于 49 株巴氏杆菌气肿亚种 Jawetz 和 Heyl 和鼠型嗜血杆菌的临床分离株中，通过与参考菌株模式的比较可以对这些分离株进行鉴定。扩增片段的序列分析显示，除“流感嗜血杆菌 murium”外，所有物种均具有较大的 ITS(ile+ala)，其中包含 tRNA(Ile(GAU))和 tRNA(Ala(UGC))基因，以及较小的 ITS(glu)，其中包含 tRNA(Glu(UUC))基因。“流感嗜血杆菌 murium”分别显示了较大和较小 ITS 片段的两个各自的 ITS 片段。在测试的巴氏杆菌气肿亚种 Jawetz 和 Heyl 和鼠型嗜血杆菌菌株中，每种 ITS 类型的长度和序列都高度保守。相反，ITS 序列显示出明显的种间差异，ITS(ile+ala)的同一性水平为 61.2%至 89.5%，ITS(glu)的同一性水平为 56.5%至 86.8%。鉴定出具有显著种间差异但在种内高度保守的 ITS 序列区域，可用于设计用于鉴定啮齿动物巴斯德氏菌科的探针，以达到种水平。
