Desai M, Linton D, Owen R J, Cameron H, Stanley J
National Collection of Type Cultures, Central Public Health Laboratory, London.
J Appl Bacteriol. 1993 Dec;75(6):574-82. doi: 10.1111/j.1365-2672.1993.tb01597.x.
DNA probes are described which identify group and fingerprint strains of the human gastric pathogen Helicobacter pylori, on the basis of well-defined band homologies. A 544 bp internal fragment of the 16S ribosomal RNA gene was generated by polymerase chain reaction (PCR) with primers derived from the Escherichia coli rRNA gene sequence. In genomic Southern blots this probe detected restriction site variation around these loci, generating simple but strain-specific molecular fingerprints. A small conserved chromosomal fragment of 1.2 kbp, Hps, species-specific for H. pylori, was obtained by cloning random HindIII fragments into pUC19. It was useful for dot-blot identification, and also separated isolates into one major and two minor groups. When results for these two probes were combined, a baseline characterization of genotype was obtained. A band-matching database of molecular fingerprints for the type strain and 63 clinical isolates of H. pylori from asymptomatic, ulcer and gastritis contexts is presented. No significant association between the genotypes at this level of definition and the associated clinical symptomatology of the isolates was detected.
描述了基于明确的条带同源性来鉴定人类胃部病原体幽门螺杆菌的菌群和指纹菌株的DNA探针。通过聚合酶链反应(PCR),利用源自大肠杆菌rRNA基因序列的引物,生成了16S核糖体RNA基因的一个544 bp内部片段。在基因组Southern印迹中,该探针检测到这些位点周围的限制性酶切位点变异,产生简单但菌株特异性的分子指纹。通过将随机的HindIII片段克隆到pUC19中,获得了一个1.2 kbp的小保守染色体片段Hps,它对幽门螺杆菌具有种特异性。它可用于斑点印迹鉴定,还可将分离株分为一个主要组和两个次要组。当将这两种探针的结果结合起来时,就获得了基因型的基线特征。给出了幽门螺杆菌模式菌株以及来自无症状、溃疡和胃炎患者的63株临床分离株的分子指纹条带匹配数据库。在这个定义水平上,未检测到基因型与分离株相关临床症状之间的显著关联。