title: Polymerase chain reaction-restriction fragment length polymorphism method for differentiation of uropathogenic specific protein gene types.
abstract: The putative pathogenicity island (PAI) containing the uropathogenic specific protein (usp) gene and three small open reading frames (orfU1, orfU2, and orfU3) encoding 98, 97, and 96 amino acid proteins is widely distributed among uropathogenic Escherichia coli (UPEC) strains. This PAI was designated as PAIusp. Sequencing analysis of PAIusp has revealed that the usp gene can be divided into two types - uspI and uspII - based on sequence variation at the 3' terminal region and the number and position of orfUs differ from strain to strain. Based on usp gene types and orfU sequential patterns, PAIusp can be divided into four subtypes. Subtyping of PAIusp is a useful method to characterize UPEC strains. In this study, we developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method to differentiate usp gene types. This method could correctly identify the usp gene type in usp-positive UPEC strains in our laboratory.
authors.affiliation: Pathobiological and Medical Diagnostics Department, Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu, 88400, Sabah, Malaysia.
authors.forename: Yun Mei
authors.initials: YM
authors.lastname: Lai
authors.affiliation: Pathobiological and Medical Diagnostics Department, Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu, 88400, Sabah, Malaysia.
authors.forename: Myo Thura
authors.initials: MT
authors.lastname: Zaw
authors.affiliation: Department of Community Medicine, Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu, 88400, Sabah, Malaysia.
authors.forename: Shamsul Bahari
authors.initials: SB
authors.lastname: Shamsudin
authors.affiliation: Pathobiological and Medical Diagnostics Department, Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu, 88400, Sabah, Malaysia. Electronic address: zawlin@ums.edu.my.
authors.forename: Zaw
authors.initials: Z
authors.lastname: Lin
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics*; D044404:Genomic Islands / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D014552:Urinary Tract Infections / Q000382:microbiology; D056689:Uropathogenic Escherichia coli / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C446159:USP protein, E coli
keywords: Pathogenicity island; Restriction fragment length polymorphism; Urinary tract infection; Uropathogenic Escherichia coli; Uropathogenic specific protein