Molecular Study of Acinetobacter Baumannii Using 16s Rrna And Bla Oxa-51 Gene Isolated from Hospitals in Duhok-kurdistan Region, Iraq

Abstract

Background and objective: Acinetobacter baumannii is pathogenic and multiresistant bacteria that cause nosocomial infection. The aim of this study are diagnosis A. baumannii by using specific bla OXA-51 and 16SrRNA. Methods: out of 150 Acinetobacter species samples were collected from patients in Azadi teaching hospital, Duhok Emergency hospital; and burn Duhok cosmetic and burn hospital from May 2020 to August 2020. The samples were phenotypically diagnosed by bacteriological strategy. Out of 100 samples were selected and by molecular level confirmed as Acinetobacter spp. Results: isolates were revealed as small, pale, and late-lactose fermenter colonies on MacConkey agar appeared as a creamy, opaque, andnon-hemolytic colony on blood agar All suspected isolates, as A. baumannii, were growing at 44ºC. By using the genus-specific bla OXA-51 primer that produced 353 bp amplification band and 75 samples were diagnosed by PCR as A. baumannii by 16S rRNA as a specific primer and showed 150 bp amplicon. 10 samples out of the 75 resembling A. baumannii were identified as a multidrug resistant isolates by method of diffusion. Disc, A. baumannii isolates displayed a high resistance rate of 85% to azithromycin and 80% to imipenem. Moreover, amikacin, meropenem, and gentamycin, Trimethoprim-Sulfamethoxazole, and ciprofloxacin, norfloxacin showed a low level of efficacy against A. baumannii isolates with the resistant rate of 88%, 90%, 90%, and 93% respectively.

Study the Role of the Efflux Pump in Multidrug-Resistant Acinetobacter baumannii

“…In the present work, 58% of the isolates showed resistance to carbapenem (CRAB) by the Kirby-Bauer disc diffusion susceptibility testing, and among these CRAB isolates, carbapenemase activity was detected by the RAPDEC Carba NP and Meropenem+EDTA CDT tests in 88.8% and 55%, respectively. which was inconsistence with studies conducted recently in Sulaimani city [15] and in Duhok city [16], furthermore, high resistance to carbapenems reported in Iran 94% in 2020 [12], 99% in 2021 [24] and in Jordan 75.1% in 2022 [39]. To determine the role of efflux pump activity, we used EP inhibitor (CCCP) to observe the reduce in the sensitivity of the tested isolates toward meropenem, the MIC was reduced by at least fourfold in the presence of 25µg/mL CCCP in 22.5% (7/31) of all isolates and 38.8% (7/18) among CRAB isolates.…”

“…These enzymes are intrinsic and naturally exist in the chromosomal DNA of A. baumannii inherited into all strains. This fact makes it a wonderful marker successfully applied for the detection of A. baumannii at the species level [16].…”

“…Using a Bio-Rad Gel Doc XR+ imaging technique, images were captured. Agarose gel electrophoresis (Cleaver, Scientific Ltd., U.K.) was used to separate the PCR products in 1x TBE buffer at 80V for 60 minutes [16].…”

Molecular Characterisation of Carbapenemase-Producing Acinetobacter baumannii isolates from Hospitalised Patients in Iraq

“…This study found resistance to imipenem (96%) and meropenem (94%) by the disc diffusion approach. Clos to this study's findings, a previous study in Dohuk by Qader (2021) reported that 80% and 90% of isolates exhibited resistance to imipenem and meropenem, respectively. However, Smail and AL-otrachi ( 2020) found a higher rate of resistance (100%) to the above-mentioned antibiotics in a study performed in Erbil.…”