|Samah Nema Abd Al-fatlawi||Molecular Variations in FimH gene of Escherichia coli Isolated from Different Sources in Holy Kerbala Province||the master degree in biology|
In the current study, sixty urine samples were collected from women with urinary tract infection symptoms aged from 18 to 60 years from attending Maternity and Women’s Hospital (obstetrics and gynecology perceptive hospital) and primary Healthy Caring Center of Muthafeen neighborhood in Kerbala province. In addition samples were taken from different environmental places involving eight poultry, two knacker shops, four drainage water four fresh water, seven swage water during 22ed November 2017 to 28ed February 2018.
Then, the isolates were identified by traditional biochemical tests , gram stain and confirmed Api 20E examination to diagnostic bacteria under current study. Biofilm was performed by 96 well microtitere plate method, 21Clinical isolates 23 and environmental isolates were underwent to the test to measuring correlation between biofilm formation and FimH gene occurrence. Additionally, FimH gene was amplified by PCR technique.
In addition, biofilm production;14(66.6%) were strong, 7(33.30%) moderate and weak biofilm producers not exit in clinical Isolates while, 13(41%) strong, 8(34.70%) moderate and 2(8.60%) for weak in environmental isolates. PCR assay was applied, 20 isolates (95.23%) positive results for clinical isolates and 17 isolates (73.9%) positive results for environmental isolates to FimH gene while twenty two PCR product involving 11 clinical isolates and 11 environmental isolates examined by sequencing assay, DNA sequence for FimH gene with its phylogenic tree wear determined for selected isolates, comparing the clinical and environmental with each other and with the global isolates which tacking from different sources and place.
Phe1, Ile13, Asn46, Asp47, Tyr48, Ile52,Asp54, Gln133, Asn135, Asp140 and Phe142 high-resolution with 3D structure of FimH protein with their active site was determined to show properties depended on amino acid sequence.
Two types of mutation were identified, missense mutation which is carrying change in DNA sequencing that leading to altered in one amino acid(Lucien to Iso-Lucien) which found in FimH gene that belongs to clinical isolates and silent mutation, known as neutral mutation, found in FimH gene belong to environmental isolates.