Roggenkamp A, Sing A, Hornef M, Brunner U, Autenrieth I B, Heesemann J
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, Clinic Innenstadt, Ludwig Maximilians University Munich, 80336 Munich, Germany.
J Clin Microbiol. 1998 Sep;36(9):2530-4. doi: 10.1128/JCM.36.9.2530-2534.1998.
From two different specimens of a chronic prosthetic hip infection taken at an interval of 2 months a slow-growing gram-negative bacterium was isolated in pure culture. The strain grew with the typical features of a small-colony variant (SCV). 16S rRNA sequencing identified the bacterium as Escherichia coli. Biochemical characterization demonstrated multiple phenotypic alterations of a mutant carrying a defect in the heme biosynthetic pathway (Hem-): (i) catalase and nitrate reductase reactions were both negative, (ii) a negative benzidine reaction demonstrated the lack of heme-containing cytochromes, and (iii) growth stimulation under anaerobic conditions as well as gentamicin resistance indicated defective aerobic respiration. PCR and Southern hybridization demonstrated that the mutation of the SCV of E. coli was localized in the hemB gene and was most likely due to a deletion of the hemB gene. On blood agar plates revertants were recognized growing as normal-sized colonies between the dominant small colonies of the strain. Feeding experiments indicated that the revertants but not the small colonies were permeable for hemin. A strong antibody response against the infecting SCV of E. coli was found. To our knowledge, this is the first report of a Hem- E. coli strain as the etiological agent of a chronic bacterial infection.
从间隔2个月采集的两份慢性人工髋关节感染标本中,分离出一株生长缓慢的革兰氏阴性菌,为纯培养物。该菌株具有小菌落变体(SCV)的典型生长特征。16S rRNA测序鉴定该细菌为大肠杆菌。生化特性表明,一株携带血红素生物合成途径缺陷(Hem-)的突变体存在多种表型改变:(i)过氧化氢酶和硝酸盐还原酶反应均为阴性;(ii)联苯胺反应阴性表明缺乏含血红素的细胞色素;(iii)厌氧条件下的生长刺激以及对庆大霉素的耐药性表明有氧呼吸存在缺陷。PCR和Southern杂交表明,大肠杆菌SCV的突变位于hemB基因,很可能是由于hemB基因缺失所致。在血琼脂平板上,回复突变体被识别为在该菌株占主导的小菌落之间生长的正常大小菌落。喂养实验表明,回复突变体而非小菌落对血红素具有通透性。发现针对感染的大肠杆菌SCV有强烈的抗体反应。据我们所知,这是关于Hem-大肠杆菌菌株作为慢性细菌感染病原体的首次报道。