Kao J S, Stucker D M, Warren J W, Mobley H L
Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore 21201, USA.
Infect Immun. 1997 Jul;65(7):2812-20. doi: 10.1128/iai.65.7.2812-2820.1997.
Urinary tract infection is the most frequently diagnosed kidney and urologic disease, and Escherichia coli is by far the most common etiologic agent. Defined blocks of DNA termed pathogenicity islands have been found in uropathogenic strains to carry genes not generally found in fecal strains. We have identified one of these regions of DNA within the chromosome of the highly virulent E. coli CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. This strain, which is cytotoxic for cultured renal cells and causes acute pyelonephritis in transurethrally infected CBA mice, contains two distinct copies of the pap operon and is hemolytic. One pap operon was localized on a cosmid clone which was used to identify three overlapping cosmid clones. By using restriction mapping, DNA hybridization, sequencing, and PCR amplification, a region of approximately 50 kb was found to be present in this uropathogenic strain and to have no corresponding sequences in E. coli K-12. This gene block also carries hemolysin genes hlyCABD. The pathogenicity island begins 7 bp downstream of dadX (catabolic alanine racemase; 26.55 min) and ends at a position in the K-12 genome 75 bp downstream of the metV tRNA gene (62.74 min); this suggests that a chromosomal rearrangement has occurred relative to the K-12 linkage map. The junctions of the pathogenicity island were verified by PCR amplification directly from the genomic DNA of strain CFT073. DNA sequencing within the boundaries of the junctions revealed genes not previously identified in E. coli or in some cases bearing no known homologs. When used as probes for DNA hybridization, these sequences were found significantly more often in strains associated with the clinical syndromes of cystitis (82%) and acute pyelonephritis (79%) than in fecal strains (19%; P < 0.001).
尿路感染是最常被诊断出的肾脏和泌尿系统疾病,而大肠杆菌是迄今为止最常见的病原体。在尿路致病性菌株中发现了被称为致病岛的特定DNA片段,这些片段携带了通常在粪便菌株中不存在的基因。我们在从一名患有急性肾盂肾炎的女性血液和尿液中分离出的高毒力大肠杆菌CFT073的染色体中鉴定出了其中一个DNA区域。该菌株对培养的肾细胞具有细胞毒性,并在经尿道感染的CBA小鼠中引起急性肾盂肾炎,它含有两个不同的pap操纵子拷贝,并且具有溶血能力。一个pap操纵子位于一个黏粒克隆上,该黏粒克隆被用于鉴定三个重叠的黏粒克隆。通过限制性图谱分析、DNA杂交、测序和PCR扩增,发现这个尿路致病性菌株中存在一个约50 kb的区域,而大肠杆菌K-12中没有相应序列。这个基因片段还携带溶血素基因hlyCABD。致病岛起始于dadX(分解代谢丙氨酸消旋酶;26.55分钟)下游7 bp处,终止于K-12基因组中metV tRNA基因(62.74分钟)下游75 bp处;这表明相对于K-12连锁图谱发生了染色体重排。致病岛的连接处通过直接从菌株CFT073的基因组DNA进行PCR扩增得到验证。连接处边界内的DNA测序揭示了以前在大肠杆菌中未鉴定出的基因,或者在某些情况下没有已知同源物的基因。当用作DNA杂交探针时,这些序列在与膀胱炎(82%)和急性肾盂肾炎(79%)临床综合征相关的菌株中出现的频率明显高于粪便菌株(19%;P < 0.001)。