Sulphonamides resistant strains are highly prevalent in uropathogenic Escherichia coli (UPEC) isolates. Sul genes encode sulphonamide resistance and are present on transferrable plasmids. Integrons (IGNs) are genetic elements containing integrase gene, attl site and gene cassettes which carry multiple antibiotic resistant genes. Class 1 integrons have been extensively studied because these were most prevalent among clinical isolates. In this study, UPEC isolates were determined for the antibiotic susceptibility patterns to four antibiotics commonly used for urinary tract infections, which include co-trimethoxazole (TMP-STX). Distribution of sul genes and integrase1 gene (intI1) was studied in TMP-STX resistant UPEC isolates by using multiplex polymerase chain reaction (mPCR). Sul genes variants were investigated by DNA sequencing of the whole open reading frame of sul1 and sul2 genes and PCR product of sul3 gene. Sul1, sul2 and sul3 genes were prevalent in 37 (24.7%) of 150 UPEC isolates. IntI1 is positive in 22 sul genes positive isolates. Of six isolates positive with sul2 genes, sul2(a) and sul2(b) variants, which were described in the previous study, in the four isolates and the two isolates respectively were observed. This is the first mPCR which investigates the prevalence of three sul genes and intI1 in the UPEC clinical isolates from two hospitals of Sabah.Asian J. Med. Biol. Res. June 2016, 2(2): 213-220

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.

Introduction: Uropathogenic virulence factors have been identified by comparing the prevalence of these among urinary tract isolates and environmental strains. The uropathogenic-specific protein (USP) gene is present on the pathogenicity island (PAI) of uropathogenic Escherichia coli (UPEC) and, depending on its two diverse gene types and the sequential patterns of three open reading frame units (orfUs) following it, there is a method to characterize UPEC epidemiologically called PAIusp subtyping. Methodology: A total of 162 UPEC isolates from Sabah, Malaysia, were tested for the presence of the usp gene and the sequential patterns of three orfUs following it using polymerase chain reaction (PCR). In addition, by means of triplex PCR, the prevalence of the usp gene was compared with other two VFs of UPEC, namely alpha hemolysin (α-hly) and cytotoxic necrotizing factor (cnf-1) genes encoding two toxins. Results: The results showed that the usp gene was found in 78.40% of UPEC isolates, indicating that its prevalence was comparable to that found in a previous study in Japan. The two or three orfUs were also associated with the usp gene in this study. All the PAIusp subtypes observed in Japan were present in this study, while subtype IIa was the most common in both studies. The usp gene was observed in a higher percentage of isolates when compared with ɑ-hly and cnf-1 genes. Conclusions: The findings in Japan and Sabah, East Malaysia, were similar, indicating that PAIusp subtyping is applicable to the characterization of UPEC strains epidemiologically elsewhere in the world.

Escherichia coli sequence type 131 (ST131) carries multiple drug resistance (MDR) genes as well as virulence genes. Drug resistant characteristics give a management problem to health care personnel. Four MDR Escherichia coli ST131 H30-Rx subclones were identified among 80 Uropathogenic E. coli (UPEC) isolates by using 4 allelic-specific Polymerase Chain Reactions (PCR) in two hospitals of Kota Kinabalu, Sabah, Malaysia.There is emergence of multidrug resistant E. coli in Kota Kinabalu.Keywords: Uropathogenic E. coli, ST131 H30-Rx subclones, multiple drug resistance, hospitals of Kota Kinabalu

The three plasmid borne alternative dihydropteroate synthetase (DHPS) genes namely sul1, sul2 and sul3 genes were heterologous in amino acid sequence and have about 40 – 50% identity. However, they have same DHPS activity with disc diffusion zone size of 6 mm with suphamethoxazole disc in our previous study. Sul1, sul2 and sul3 genes were observed in sulphamethoxazole resistant uropathogenic Escherichia coli (UPEC). In this study, all the three genes were cloned into E. coli host and minimum inhibitory concentration (MIC) was investigated for each sul gene to compare the activities of sul genes. The MIC values of E.coli containing sul2, sul1 and sul3 genes inserted recombinant plasmid were observed to have 18.5 mg/ml, 18 mg/ml and 17.5 mg/ml respectively as mean value of five experimental results. Although comparable MICs were obtained as a result, the MIC value was highest in E. coli carrying sul2 gene indicating that this DHPS enzyme activity of sul2 was strongest among three sul genes.