Molecular Analysis of Acinetobacter baumannii Strains Isolated in Lebanon Using Four Different Typing Methods (original) (raw)
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Infections caused by Carbapenem-Resistant Acinetobacter baumannii (CRAB) have become an essential healthcare-associated problem. We used whole-genome sequencing (WGS) to characterize A. baumannii collected from a hospital in Lebanon. The genotypic relatedness was assessed by pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), and whole-genome SNP-based phylogenetic analysis (wg-SNP). A. baumannii PCR-based replicon typing (AB-PBRT) was also used to analyze the plasmid content. PFGE demonstrated that most isolates that belonged to a unique clonal type were assigned to ST2 of the international clone II. Phenotypic antimicrobial susceptibility testing revealed that the isolates were extensively drug-resistant (XDR). Intrinsic β-lactam resistance genes, including blaADC25 (Acinetobacter-Derived Cephalosporinase) and the blaOXA66 (a variant of blaOXA51), were among the common resistance determinants. AB-PBRT, which categorizes A. baumannii plasmids into homology ...
Antimicrobial Resistance & Infection Control
Background Antibiotic use is largely under-regulated in Egypt leading to the emergence of resistant isolates. Carbapenems are last resort agents to treat Acinetobacter baumannii infections resistant to other classes of antibiotics. However, carbapenem-resistant isolates are emerging at an alarming rate. This study aimed at phenotypically and molecularly characterizing seventy four carbapenem-unsusceptible A. baumannii isolates from Egypt to detect the different enzymes responsible for carbapenem resistance. Methods Carbapenemase production was assessed by a number of phenotypic methods: modified Hodge test (MHT), carbapenem inactivation method (CIM), combined disc test (CDT), CarbAcineto NP test and boronic acid disc test. Polymerase chain reaction (PCR) was used to screen the isolates for the presence of some genes responsible for resistance to carbapenems, as well as some insertion sequences. Results PCR amplification of class D carbapenemases revealed the prevalence of blaOXA-51 ...
Infectious Disorders - Drug Targets, 2019
Background:Acinetobacter baumannii is an opportunistic pathogen, which causes a wide range of infections in hospitals, especially in intensive care units. Nowadays, due to the high resistance of Acinetobacter bumanni to antibiotics, this study, in addition to the phenotypic and genotypic investigations of drug resistance, focused on determining the molecular types of Acinetobacter baumannii isolated from patients in Khorramabad city by the pulsed-field gel electrophoresis (PFGE) method.Materials and Methods:In this cross-sectional study, 50 samples of Acinetobacter baumannii were collected from educational hospitals in Khorramabad city, Iran, from January to August 2015. They were identified in the laboratory using biochemical tests and culture methods. After determining the drug resistance pattern by the disc diffusion method and percentage of resistance genes to carbapenems, Acinetobacter baumannii isolates were analyzed using the PFGE method using the Apa1 enzyme.Results:The high...
2015
It has previously been shown that carbapenem-resistant Acinetobacter baumannii are frequently detected in Saudi Arabia. The present study aimed to identify the epidemiology and distribution of antibiotic resistance determinants in these bacteria. A total of 83 A. baumannii isolates were typed by PFGE, and screened by PCR for carbapenemase genes and insertion sequences. Antibiotic sensitivity to imipenem, meropenem, tigecycline and colistin were determined. Eight different PFGE groups were identified, and were spread across multiple hospitals. Many of the PFGE groups contained isolates belonging to World-wide clone 2 (WW2). Carbapenem resistance or intermediate resistance was detected in 69% of isolates. The blaVIM gene was detected in 94% of isolates, while blaOXA-23-like genes were detected in 58%. The data demonstrate the co-existence and wide distribution of a number of clones of carbapenem-resistant A. baumannii carrying multiple carbapenem-resistance determinants within hospitals in the Eastern Region of Saudi Arabia.
Antibiotics
At the Bundeswehr Hospitals of Hamburg and Westerstede, patients repatriated from subtropical war and crisis zones of Northern Africa and the Middle East were medically treated, including microbiological assessment. Within a six-year interval, 16 Acinetobacter spp. strains, including 14 Acinetobacter baumannii (Ab) isolates with resistance against carbapenems and origins in Afghanistan (n = 4), Iraq (n = 2), Libya (n = 2), and Syria (n = 8) were collected. While clonal relationships of Libyan and Syrian strains had been assessed by superficial next generation sequencing (NGS) and “DiversiLab” repetitive elements sequence-based (rep-)PCR so far, this study provides core genome-based sequence typing and thus more detailed epidemiological information. In detail, sequencing allowed a definitive species identification and comparison with international outbreak-associated Ab strains by core genome multi locus sequence typing (cgMLST) and the identification of MLST lineages, as well as the...
Japanese Journal of Infectious Diseases, 2021
We aimed to investigate the clonal relationships, common sequence types, and carbapenemase genes in 177 non-repetitive blood culture isolates of Acinetobacter baumannii collected from patients at three university hospitals in Turkey in 2016. Molecular epidemiological characteristics of the isolates were examined using pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) (Pasteur scheme-cpn60, fusA, gltA, pyrG, recA, rplB, and rpoB). Multiplex PCR was used to investigate the carbapenemase genes, including bla OXA-23-like , bla OXA-24-like , bla OXA-48-like , bla OXA-58-like , bla IMP , bla VIM , and bla NDM. PFGE genotyping yielded 92 pulsotypes with a clustering ratio of 69.7%. As per a ≥85% similarity coefficient, 159 (90.9%) isolates were found to be clonally related. The bla OXA-23-like and bla OXA-58-like genes were identified in 100% and 28.2% of the isolates, respectively. The bla NDM gene was identified in two isolates. The MLST analysis included 54 isolates with different pulsotypes, and 29 sequence types (STs). Most of the isolates (n = 36) belonged to the clonal complex (CC)2, one isolate belonged to CC1, and one isolate belonged to CC164. Sixteen new STs (ST1235-ST1250) were identified. Identifying both global ST2 and a large number of new STs, revealed high genetic diversity in A. baumannii isolates in the study population.
Journal of clinical microbiology, 2015
The molecular epidemiology and mechanisms of resistance of carbapenem-resistant Acinetobacter baumannii (CRAB) were determined in hospitals in the states of the Cooperation Council for the Arab States of the Gulf (Gulf Cooperation Council [GCC]), namely, Saudi Arabia, United Arab Emirates, Oman, Qatar, Bahrain, and Kuwait. Isolates were subjected to PCR-based detection of antibiotic resistance genes and repetitive sequence-based PCR (rep-PCR) assessments of clonality. Selected isolates were subjected to multilocus sequence typing (MLST). We investigated 117 isolates resistant to carbapenem antibiotics (either imipenem or meropenem). All isolates were positive for OXA-51. The most common carbapenemases were the OXA-23-type, found in 107 isolates, followed by OXA-40-type (OXA-24-type), found in 5 isolates; 3 isolates carried the ISAba1 element upstream of blaOXA-51-type. No OXA-58-type, NDM-type, VIM-type, or IMP-type producers were detected. Multiple clones were detected with 16 clus...
2015
BackgroundA. baumannii has emerged as an important nosocomial pathogen with an outstanding ability to acquire multidrug resistant mechanisms. In this study, we investigate the molecular epidemiology and carbapenem resistance mechanisms of A. baumannii in Tripoli, Northern Lebanon.MethodsOne hundred sixteen non-duplicate isolates isolated between 2011 and 2013 in different hospitals in Tripoli, Lebanon from Lebanese patients and wounded Syrian patients during Syrian war were studied. Antibiotic susceptibility testing was determined by agar disc diffusion and Etest. Carbapenemase-encoding genes were investigated by PCR. All isolates were typed by blaOXA-51-like sequence based typing (SBT) and 57 isolates were also analysed by MLST using Pasteur’s scheme followed by eBURST analysis.ResultsOf the 116 isolates, 70 (60 %) showed a carbapenem resistance phenotype. The blaOXA-23 with an upstream insertion of ISAba1 was the major carbapenem resistance mechanism and detected in 65 isolates. F...
Genotyping of carbapenem resistant Acinetobacter baumannii isolated from Egyptian patients
Novel Research in Microbiology Journal
Acinetobacter baumannii has recently been known as a major cause of hospital-and community-acquired infections. Carbapenem resistant A. baumanni (CRAB) has been recorded to be resistant to nearly all antibiotics, including the last resort antibiotics; carbapenems. This study aimed to detect the carbapenem resistance levels and mechanisms, in addition to the genotyping of A. baumanni in Upper Egypt. About 200 clinical samples were collected from different wards of Sohag University Hospital, Egypt, from which 20 A. baumannii isolates were recovered and then identified using conventional methods and Polymerase Chain Reaction (PCR). Antibiotic sensitivity testing was carried out using the Disk diffusion method, followed by PCR testing of the common carbapenemase-encoding genes, including OXA-51, OXA-58, KPC, GES, IMP, NDM, VIM, SIM, and GIM. Genotyping was performed using the Enterobacterial Repetitive Intergenic Consensus-Polymerase chain reaction (ERIC-PCR). About 85 % of A. baumannii strains were multidrug resistant (MDR), and high rate of Extreme drug resistant (XDR) A. baumannii (70 %) was detected. Carbapenem resistance was detected in 65 % of A. baumannii isolates, 70.58 % of MDR isolates, and 85.7 % of XDR isolates, respectively. Carbapenemase-encoding genes, including bla OXA-51 , VIM, NDM, and GES, were detected in 100 %, 100 %, 76.9 2 % and 76.92 % of the carbapenem resistant A. baumannii (CRAB) isolates, respectively. The bla IMP and bla KPC genes had lower prevalence rates of 15.38 % and 30.77 %; respectively, whereas the SIM, GIM, and OXA-58 genes were not detected in any of the tested A. baumanni isolates. All of the MDR isolates carried three or more the carbapenemases encoding genes, and 85.7 % of the XDR isolates carried four or more of the carbapenemase-encoding genes. The dendrogram