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Polymerase chain reaction detection of haemolysin D gene (hlyD) in uropathogenic Escherichia coli as a novel diagnostic test for urinary tract infection

[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”

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Antimicrobial susceptibility of urinary bacterial isolates of pregnant women attending antenatal clinics of selected hospitals in Ilorin, Nigeria

*1Baba, R. T., 1Adedayo, M. R., 1Awe, S., and 2Bale, M. I.

1Department of Microbiology, Faculty of Pure and Applied Sciences, Kwara State University,  Malete, Kwara State, Nigeria

2Department of Microbiology, University of Ilorin Teaching Hospital, Ilorin, Nigeria

*Correspondence to: omotayo.rasheedat@gmail.com; 07065391534: 09026144501

                         

Abstract:

 Background: Urinary tract infection (UTI) in pregnancy is associated with significant morbidity for both mother and baby. This study was aimed at determining the urinary bacterial isolates and their susceptibility to selected antibiotics among pregnant women attending antenatal clinics of selected Hospitals in Ilorin, Nigeria.

Methodology: A total of 300 pregnant women between the ages of 15 and 44 years were selected by random sampling technique from antenatal clinics of three randomly selected healthcare facilities in Ilorin, Nigeria; Civil Service Hospital, Sobi Specialist Hospital and Okelele Primary Health Center, between July and October 2021.  Clean-catch mid-stream specimens of voided urine were collected from each participant, cultured on CysteineLactose-Electrolyte Deficient (CLED) and Blood agar plates, and incubated aerobically at 37oC for 24 hours. The presence of significant bacteriuria (≥105 CFU/ml) was determined on the culture plate using the plate count method. Antibiotic susceptibility testing to selected antibiotics was done using Kirby-Bauer disk diffusion technique. Cefoxitin (30𝜇g) was used as surrogate to determine phenotypic methicillin resistance in staphylococcus isolates, and the methicillin resistance (mecA) gene was detected by conventional PCR assay. Continue reading “Antimicrobial susceptibility of urinary bacterial isolates of pregnant women attending antenatal clinics of selected hospitals in Ilorin, Nigeria”