Guyer D M, Kao J S, Mobley H L
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
Infect Immun. 1998 Sep;66(9):4411-7. doi: 10.1128/IAI.66.9.4411-4417.1998.
Urinary tract infection is the most frequently diagnosed kidney and urologic disease and Escherichia coli is by far the most common etiologic agent. Uropathogenic strains have been shown to contain blocks of DNA termed pathogenicity islands (PAIs) which contribute to their virulence. We have defined one of these regions of DNA within the chromosome of a highly virulent E. coli strain, CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. The 57,988-bp stretch of DNA has characteristics which define PAIs, including a size greater than 30 kb, the presence of insertion sequences, distinct segmentation of K-12 and J96 origin, GC content (42.9%) different from that of total genomic DNA (50.8%), and the presence of virulence genes (hly and pap). Within this region, we have identified 44 open reading frames; of these 44, 10 are homologous to entries in the complete K-12 genome sequence, 4 are nearly identical to the sequences of E. coli J96 encoding the HlyA hemolysin, 11 encode P fimbriae, and 19 show no homology to J96 or K-12 entries. To determine whether sequences found within the junctions of the PAI of CFT073 were common to other uropathogenic strains of E. coli, 11 probes were isolated along the length of the PAI and were hybridized to dot blots of genomic DNA isolated from clinical isolates (67 from patients with acute pyelonephritis, 38 from patients with cystitis, 49 from patients with catheter-associated bacteriuria, and 27 from fecal samples). These sequences were found significantly more often in strains associated with the clinical syndromes of acute pyelonephritis (79%) and cystitis (82%) than in those associated with catheter-associated bacteriuria (58%) and in fecal strains (22%) (P < 0.001). From these regions, we have identified a putative iron transport system and genes other than hly and pap that may contribute to the virulent phenotype of uropathogenic E. coli strains.
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。已证明尿路致病性菌株含有被称为致病岛(PAIs)的DNA片段,这些片段有助于其毒力。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菌株CFT073的染色体中定义了其中一个DNA区域。这个57988碱基对的DNA片段具有定义致病岛的特征,包括大小大于30kb、存在插入序列、与K-12和J96来源的明显分割、GC含量(42.9%)与总基因组DNA(50.8%)不同,以及存在毒力基因(hly和pap)。在这个区域内,我们鉴定出44个开放阅读框;在这44个中,10个与完整的K-12基因组序列中的条目同源,4个与编码HlyA溶血素的大肠杆菌J96序列几乎相同,11个编码P菌毛,19个与J96或K-12条目没有同源性。为了确定在CFT073致病岛连接处发现的序列是否为其他尿路致病性大肠杆菌菌株所共有,沿着致病岛的长度分离出11个探针,并与从临床分离株(67株来自急性肾盂肾炎患者,38株来自膀胱炎患者,49株来自导管相关性菌尿患者,27株来自粪便样本)分离的基因组DNA的斑点杂交膜进行杂交。这些序列在与急性肾盂肾炎(79%)和膀胱炎(82%)临床综合征相关的菌株中比在与导管相关性菌尿(58%)和粪便菌株(22%)相关的菌株中出现的频率明显更高(P<0.001)。从这些区域中,我们鉴定出一个假定的铁转运系统以及除hly和pap之外的其他可能有助于尿路致病性大肠杆菌菌株毒力表型的基因。