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间质性膀胱炎及对照患者膀胱活检组织中细菌16S rRNA基因的聚合酶链反应扩增

Polymerase chain reaction amplification of bacterial 16S rRNA genes in interstitial cystitis and control patient bladder biopsies.

作者信息

Keay S, Zhang C O, Baldwin B R, Jacobs S C, Warren J W

机构信息

Department of Medicine, University of Maryland School of Medicine, Baltimore, USA.

出版信息

J Urol. 1998 Jan;159(1):280-3. doi: 10.1016/s0022-5347(01)64082-5.

Abstract

PURPOSE

Several characteristics of the chronic bladder disease called interstitial cystitis (IC) suggest an infectious etiology. However, a single causative organism has not been convincingly cultured in vitro, and DNA for a variety of microorganisms has been found inconsistently in bladder biopsies from IC patients. We therefore looked for a possible bacterial cause for IC by using a sensitive nested PCR assay on cystoscopic bladder biopsy specimens obtained from IC patients and controls.

MATERIALS AND METHODS

Bladder biopsies were obtained at cystoscopy from 6 IC patients and 6 controls. DNA was extracted from these specimens and PCR with 2-round amplification performed using nested primers from a highly conserved region of the bacterial 16s rRNA gene. Amplified DNA was purified and sequenced using the Sequenase PCR Product Sequencing Kit, and the sequences obtained were compared with bacterial rRNA gene sequences recorded in GenBank.

RESULTS

Biopsy specimens from all 6 patients and 6 controls were positive by PCR for DNA encoding bacterial 16s rRNA. Sequence data indicated a predominant microorganism in 10 of the 12 specimens, with > 95% homology to DNA from several different genera of bacteria including Acinetobacter, Propionobacterium, Salmonella, and Escherichia. None of the organisms identified by PCR had been cultured from tissue or urine obtained simultaneously from these persons, using sensitive culture techniques.

CONCLUSIONS

These data indicate no difference between IC patients and controls in the proportion of bladder biopsies with PCR positivity or the type(s) of organism present, providing additional evidence that IC is not associated with infection by a particular type of bacterium.

摘要

目的

称为间质性膀胱炎(IC)的慢性膀胱疾病的几个特征提示其病因与感染有关。然而,尚未在体外令人信服地培养出单一的致病生物体,并且在IC患者的膀胱活检组织中不一致地发现了多种微生物的DNA。因此,我们通过对从IC患者和对照组获得的膀胱镜检查膀胱活检标本进行敏感的巢式PCR检测,寻找IC可能的细菌病因。

材料与方法

在膀胱镜检查时从6例IC患者和6例对照中获取膀胱活检组织。从这些标本中提取DNA,并使用来自细菌16s rRNA基因高度保守区域的巢式引物进行两轮扩增的PCR。扩增的DNA经纯化后使用Sequenase PCR产物测序试剂盒进行测序,并将获得的序列与GenBank中记录的细菌rRNA基因序列进行比较。

结果

所有6例患者和6例对照的活检标本经PCR检测编码细菌16s rRNA的DNA均呈阳性。序列数据表明,12个标本中有10个存在主要微生物,与包括不动杆菌属、丙酸杆菌属、沙门氏菌属和大肠杆菌属在内的几种不同细菌属的DNA具有> 95%的同源性。使用敏感培养技术,通过PCR鉴定出的生物体均未从这些人同时获得的组织或尿液中培养出来。

结论

这些数据表明,IC患者和对照组之间膀胱活检标本PCR阳性比例或存在的生物体类型没有差异,这进一步证明IC与特定类型细菌的感染无关。

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