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荧光原位杂交技术快速检测尿路感染样本中的三种不同致病菌。

Fluorescence in situ hybridization rapidly detects three different pathogenic bacteria in urinary tract infection samples.

机构信息

Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Jiefang Road 238, Wuhan, 430060, Hubei Province, China.

出版信息

J Microbiol Methods. 2010 Nov;83(2):175-8. doi: 10.1016/j.mimet.2010.08.015. Epub 2010 Aug 31.

Abstract

The detection of pathogenic bacteria in urine is an important criterion for diagnosing urinary tract infections (UTIs). By using fluorescence in situ hybridization (FISH) with rRNA-targeted, fluorescently labeled oligonucleotide probes, bacterial pathogens present in urine samples were identified within 3-4 h. In this study, three probes that are specific for Escherichia coli, Enterococcus faecalis and Staphylococcus aureus were designed based on the conserved 16S RNA sequences, whereas probe Eub338 broadly recognizes all bacteria. We collected a total of 1000 urine samples, and 325 of these samples tested positive for a UTI via traditional culturing techniques; additionally, all 325 of these samples tested positive with the Eub338 probe in FISH analysis. FISH analyses with species-specific probes were performed in parallel to the test the ability to differentiate among several pathogenic bacteria. The samples for these experiments included 76 E. coli infected samples, 32 E. faecalis infected samples and 9 S. aureus infected samples. Compared to conventional methods of bacterial identification, the FISH method produced positive results for >90% of the samples tested. FISH has the potential to become an extremely useful diagnostic tool for UTIs because it has a quick turnaround time and high accuracy.

摘要

尿液中病原体的检测是诊断尿路感染(UTI)的重要标准。通过使用针对 rRNA 的荧光原位杂交(FISH)技术和荧光标记的寡核苷酸探针,可在 3-4 小时内鉴定尿液样本中的细菌病原体。在这项研究中,根据保守的 16S RNA 序列设计了三种针对大肠杆菌、粪肠球菌和金黄色葡萄球菌的特异性探针,而探针 Eub338 则广泛识别所有细菌。我们共收集了 1000 份尿液样本,其中 325 份通过传统培养技术检测出 UTI 呈阳性;此外,所有 325 份样本在 FISH 分析中均与 Eub338 探针呈阳性。通过平行进行种特异性探针的 FISH 分析来测试区分几种致病细菌的能力。这些实验的样本包括 76 份大肠杆菌感染样本、32 份粪肠球菌感染样本和 9 份金黄色葡萄球菌感染样本。与传统的细菌鉴定方法相比,FISH 方法对检测的样本产生了 90%以上的阳性结果。FISH 有可能成为一种非常有用的 UTI 诊断工具,因为它具有快速周转时间和高准确性。

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