Molecular Detection of Some Antibiotic Resistance Genes of Acinetobacter baumannii Isolates from Patients in Erbil/KRG-Iraq

Hamad, Hazheer Mawlood (2023) Molecular Detection of Some Antibiotic Resistance Genes of Acinetobacter baumannii Isolates from Patients in Erbil/KRG-Iraq. Masters thesis, Koya University.

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Abstract

Recently, multidrug-resistant Acinetobacter baumannii has raised its impact on patient care specifically after covid-19 pandemic. The potential revolution of Acinetobacter baumannii is driven by the undetectable numbers of gene resistance that is acquired. Of 570 patient specimens including blood, sputum, urine, stool, pus, swab, and body fluid, 23 (4.04%) Acinetobacter baumannii were identified by biochemical tests and VITEK 2 compact system. Identification of Acinetobacter baumannii has been studied phenotypically by conventional biochemical tests and VITEK 2 compact system, and genotypically by using polymer chain reaction (PCR) to detect 16S-23S rRNA gene intergenic spacer region (ITS) and sequencing the amplicons by Sanger sequence technique. Isolates characterizations included biochemical identification, plasmid detection, quantification of biofilm formation using microtiter plate method, antibiotic resistance profile by using VITEK 2 compact system, and PCR amplification gene resistance. Twenty-two resistance genes were targeted including β- lactamase genes: blaOXA-51, blaOXA-58, blaOXA-23, blaTEM, blaNDM, and blaSHV, aminoglycoside genes: aph(3’)-VI, aacA4, aadB, strA and strB, sulfonamide genes: sul I and sul II , multidrug efflux gene adeB , tetracycline genes: tetA and tetB, macrolide genes: msr(E), mph(E) and erm 42, Trimethoprim gene: dhfr1 and Fluoroquinolones genes: parC and gyrA. Out of 100 (21.28%) who were covid-19 patients, 13 (13%) isolates were Acinetobacter baumannii. All Acinetobacter baumannii isolates carried a plasmid and there were no strong biofilm formation. morever, 41.2% of the isolates formed medium biofilm which is statistically significant, 35.3% of the isolates formed weak biofilm and 17.6% of the isolates not formed biofilm which are statistically non-significant. Both bacterial chromosome and plasmid carried resistance genes and each isolate with at least seven resistance genes. Besides, this is the first study to display the genetic resistance epidemics in Erbil/Iraq, blaTEM detected the disseminated gene in isolates, and aadB was not encoded by isolates. The majority of the resistance genes that were not detected by the isolate chromosome were detected on the plasmid of the isolate. Furthermore, new carbapenemase gene sequences have been identified to be acquired by plasmids from other bacterial genera such as Klebsiella pneumonia and Escherichia coli which had coinfected cases with Acinetobacter baumannii in the study. The new genes are blaNDM, (Acinetobacter baumannii strain carbapenems resistance HK19; accession number: OP572243) and blaTEM (Acinetobacter baumannii strain beta-lactam resistance HK22; accession number: OP572244). In addition, a new strain Stenotrophomones maltophilia (accession number OP422244) was submitted to GenBank and acquired new beta-lactamase genes from Acinetobacter baumannii blaNDM and blaOXA-23 genes under accession numbers OP595162, OP595163 respectively. The isolate’s metabolism pathway used was carbohydrates at 93.8% and coumarate at 100%. The results demonstrated the pattern role of plasmid in Acinetobacter baumannii resistance and the alternative metabolic pathway in all strains to survive was coumarate metabolic pathway, although the isolates metabolic differences but all strains used the coumarate pathway to survive.

Item Type:	Thesis (Masters)
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Subjects:	Q Science > QH Natural history > QH301 Biology
Divisions:	Faculty of Science and Health > Department of Biology > M.Sc. Thesis
Depositing User:	Mr. Rebwar Mohammed Jarjis
Date Deposited:	16 Nov 2023 09:02
Last Modified:	16 Nov 2023 09:02
URI:	http://eprints.koyauniversity.org/id/eprint/434

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