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首页> 外文期刊>Journal of Clinical Microbiology >Rapid and specific detection of the pap, afa, and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction.
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Rapid and specific detection of the pap, afa, and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction.

机译：通过聚合酶链反应快速，特异地检测尿路致病性大肠杆菌菌株中pap，afa和sfa粘附素编码操纵子。

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摘要

Adhesin-encoding operons (pap, sfa/foc, and afa) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. A quick and sensitive assay to identify these operons was developed by using the polymerase chain reaction (PCR). Three pairs of 25-mer primers were defined from the sequences of the DNA fragments used as probes in hybridization studies to identify each of the three operons, and the six primers were used together in a single reaction of amplification. To validate the PCR approach for detection of adhesin-encoding operons among clinical isolates, we investigated a collection of 97 E. coli isolates with the following characteristics: all isolates originated from the urine of patients with pyelonephritis, and the adhesin responsible for specific binding of the isolates to uroepithelial cells was previously characterized by phenotypic assays, as well as genotypic tests based on hybridization. There was a perfect correlation between the results obtained with the PCR approach and those previously obtained by using DNA probes. These results indicate that the PCR method, which is highly specific and easier to perform than the hybridization method, is a powerful genotypic assay for detection of adhesin-encoding operons. Thus, this assay can be recommended for clinical use to detect virulent urinary E. coli strains, as well as for epidemiological studies.
机译：粘附素编码操纵子（pap，sfa / foc和afa）在与尿路感染相关的大肠杆菌菌株中普遍存在。通过使用聚合酶链反应（PCR），开发了一种快速灵敏的测定法来鉴定这些操纵子。从杂交研究中用作探针的DNA片段序列中定义了三对25-mer引物，以鉴定三个操纵子中的每一个，并将这六个引物一起用于单个扩增反应中。为了验证用于检测临床分离株中粘附素编码操纵子的PCR方法，我们调查了具有以下特征的97种大肠杆菌分离株的集合：所有分离株均来自肾盂肾炎患者的尿液，并且粘附素负责与以前通过表型分析以及基于杂交的基因型测试对尿道上皮细胞分离株进行了表征。 PCR方法获得的结果与以前使用DNA探针获得的结果之间存在完美的关联。这些结果表明，PCR方法比杂交方法具有更高的特异性和操作性，是检测粘附素编码操纵子的强大基因型检测方法。因此，该方法可推荐用于临床检测有毒的尿液大肠杆菌菌株以及流行病学研究。

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