[1]Abubakar, N. H., [2]Aliyu, M., *[3]Jibril, M., and 2Mohammed, Y.
1Kano State Secondary Schools Management Board, Kano, Nigeria
2Department of Medical Microbiology and Parasitology, Faculty of Clinical Sciences, College of Health Sciences Bayero University, Kano, Nigeria
3Department of Pharmacology and Therapeutics, Faculty of Pharmaceutical Sciences, College of Natural and Pharmaceutical Sciences, Bayero University, Kano, Nigeria
*Correspondence to: murtalamj@yahoo.com; +2348034453990; ORCID: 0000-0003-2554-5552
Abstract:
Background: Urinary tract infection (UTI) is a common and sometime serious infectious disease diagnosed using conventional urine culture as the ‘gold standard’ for identifying Escherichia coli, the most common causative agent. However, due to the slow turn-around-time and other challenges of urine culture, this study explores the use of a novel biomolecular polymerase chain reaction (PCR) approach to detect the presence of haemolysin D gene (hlyD) that encodes a unique virulence factor of uropathogenic E. coli (UPEC) for its rapid identification in UTI.
Methodology: Primers from UPEC CFT073 and non-pathogenic E. coli K-12 MG1655 strains provided by Nottingham Trent University, England, UK were used to investigate the presence of haemolysin D gene (hlyD) in UPEC. The hlyD primers were developed from hlyD with locus number C_RS01660 on UPEC CFT073 strain using the NCBI, virulence finder, and Island viewer, and used in a PCR assay to target the hlyD in UPEC. Three sets of PCR templates were designed (UPEC, E. coli, and “No template”), each with internal and external controls amplified in a multiplex PCR assay, and agarose gel electrophoresis was used to separate the amplicons, and determine the specificity of hlyD for UPEC. Continue reading “Polymerase chain reaction detection of haemolysin D gene (hlyD) in uropathogenic Escherichia coli as a novel diagnostic test for urinary tract infection”