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标题：A New Primer Set for Detection of fimH Gene in Escherichia coli Isolates
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作者：Yun Mei Lai ; Rina Norgainathai ; Myo Thura Zaw 等
期刊名称：International Journal of Collaborative Research on Internal Medicine & Public Health
电子版ISSN：1840-4529
出版年度：2015
卷号：7
期号：4
出版社：DRUNPP Sarajevo
摘要：Background: The FimH protein is the tip fibrillum of the type 1 fimbria which is adhesive organelle of Escherichia coli (E.coli). Nucleotide sequence variations of fimH gene were common among commensal E. coli, intestinal pathogenic strains and extraintestinal pathogens. Because many researchers pointed out FimH adhesin plays an important role in E.coli – mediated urinary tract infections (UTI), the researchers have tried to develop FimH - based anti–urinary tract infection vaccine. There are different prevalence rate of fimH gene in UPEC isolates ranging from 86% to 97.5%. Aim and Objectives: In this study, a sensitive, specific and reproducible primer sets for Polymerase Chain Reaction (PCR) method to detect fimH gene in uropathogenic Escherichia coli (UPEC) isolates has been designated. Methodology: To get the fimH whole gene sequence, a set of primer covering whole gene PCR was done on 10 UPEC isolates and DNA sequence of these PCR products were aligned to select conserved areas to design new primer set. Forty UPEC isolates, 12 commensal E.coli isolates, 6 environmental E.coli isolates and 4 Entero-hemorrhagic E.coli (EHEC) isolates were studied for prevalence of fimH gene by using newly designated primer set. Results: Out of these isolates 37 (92.5%) UPEC isolates, 6 (50%) commensal isolates, 5 (83%) environmental isolates and all the 4 (100%) EHEC isolates were positive for fimH gene. Conclusion: A new primer set was successfully designed by DNA sequencing for the fimH gene of UPEC isolates and nucleotide alignment analysis. This primer set will be useful for further screening of fimH gene for molecular study of E.coli isolates
关键词：uropathogenic Escherichia coli; fimH gene; polymerase chain reaction; new primer set