PCR-based DNA fingerprinting (REP-PCR, AP-PCR) and pulsed-field gel electrophoresis characterization of a nosocomial outbreak caused by imipenem- and meropenem-resistant Acinetobacter baumannii

Bou, Germán; Cerveró, G; Domínguez, M. Ángeles; Quereda, C.; Martı́nez-Beltrán, J

doi:10.1046/j.1469-0691.2000.00181.x

Cited by 121 publications

(89 citation statements)

References 35 publications

(57 reference statements)

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“…Resistant bacteria are circulated in clinical settings by clonal expansion and HGT (Koornhof et al, 2001;Juhas et al, 2009). rep-PCR has been proven to be an expedient and useful method for the epidemiological investigation in order to determine species lineages of A. baumannii nosocomial outbreaks (Bou et al, 2000;Lin et al, 2014;Misbah et al, 2004). This method has the advantage of being faster to perform than multilocus sequence typing and PFGE, despite the inter-laboratory variability.…”

Section: Discussionmentioning

confidence: 99%

“…To study the clonal lineage of the A. baumannii isolates, whole genomic DNA preparations were prepared for computer-assisted repetitive extragenic palindromic PCR (rep-PCR) typing using primer combinations rep-1 and rep-2 (Table 1) as described previously by Bou et al (2000). Banding patterns were analysed by Gel Compare II software version 4.0 (Applied Maths).…”

Section: Methodsmentioning

confidence: 99%

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Class 1 integrons in non-clonal multidrug-resistant Acinetobacter baumannii from Iran, description of the new bla IMP-55 allele in In1243

Azizi

Shakibaie

Badmasti

et al. 2016

Journal of Medical Microbiology

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Infections and outbreaks caused by multidrug-resistant Acinetobacter baumannii (MDR-AB) are prevalent and have been reported worldwide over the past 20 or more years. Class 1 integron in MDR-AB plays an important role in the spread of antibiotic resistance in clinical settings. This study has been conducted to evaluate the detection of metallo-b-lactamase, characterization of class 1 integron and determination of clonal relatedness among A. baumannii hospital isolates. Sixty-five clinical isolates of MDR-AB were recovered from two Iranian hospital's intensive care units from February to August 2013. Integrase (intI1) and bla IMP genes were detected in 70.8 % (n=46/65) and 9.23 % (n=6/65) of the isolates using PCR assay, respectively. No other metallo-b-lactamase genes (bla VIM , bla SIM and bla NDM ) were detected. PCR sequencing of integron gene cassette revealed the following arrays: bla OXA10 -aacA4-bla IMP-55 -cmlA5 (as a novel array was designated In1243), aacC1 and aadA1. Analysis of bla IMP gene revealed a new allele designated as bla . Gene transfer experiment by conjugation showed the 36 kb conjugative plasmid harbouring In1243. The clonal assessment by repetitive extragenic palindromic PCR demonstrated a high-degree relatedness among the strains, but strains harbouring In1243 displayed a different repetitive extragenic palindromic PCR profile. In this study, we found that a novel class 1 integron (In1243) that encoded a new bla IMP allele resided on a transferable plasmid in non-clonal strains of MDR-AB.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Class 1 integrons in non-clonal multidrug-resistant Acinetobacter baumannii from Iran, description of the new bla IMP-55 allele in In1243

Azizi

Shakibaie

Badmasti

et al. 2016

Journal of Medical Microbiology

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“…PCR amplification was performed in a reaction mix containing REP1 and REP2 primers (50 pmol ml 21 ), dNTPs mixture (0.2 mM), MgCl 2 (3 mM), 1 unit Taq DNA polymerase (Thermo Scientific), 1| PCR buffer and bacterial DNA template (2 ml) in a final volume of 25 ml. The following temperature profile was used: initial denaturation at 95 uC for 10 min, 30 cycles of denaturation at 95 uC for 1 min, annealing at 45 uC for 1 min, extension at 72 uC for 2 min and a final extension at 72 uC for 16 min (Bou et al, 2000). PCR products were run in 2% agarose gel and electrophoresed through TBE 0.5| buffer.…”

Section: Antibacterial Susceptibility Testingmentioning

confidence: 99%

Limited genetic diversity and extensive antimicrobial resistance in clinical isolates of Acinetobacter baumannii in north-east Iran

Farsiani

Mosavat

Soleimanpour

et al. 2015

Journal of Medical Microbiology

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This study determined the mechanisms and patterns of antimicrobial resistance among the isolates obtained from different wards of a teaching hospital in the city of Mashhad in north-east Iran. Between January 2012 and the end of June 2012, 36 isolates of Acinetobacter baumannii were collected from different wards of Ghaem Hospital. Antimicrobial susceptibility testing and epsilometer testing (E-test) were performed. The genetic resistance determinants of A, B and D classes of b-lactamases, aminoglycoside modifying enzymes (AMEs), efflux pumps and ISAba1 elements were assessed by PCR. Repetitive extragenic palindromic element (REP)-PCR was performed to find the genetic relatedness of the isolates. Colistin was the most effective antibiotic of those tested, where all isolates were susceptible. E-test results revealed high rates of resistance to imipenem, ceftazidime and ciprofloxacin. The majority of isolates (97 %) were multidrug-resistant. OXA-51, OXA-23 and tetB genes were detected in all isolates, but OXA-58, IMP and tetA were not detected. The prevalence of OXA-24, bla TEM , bla ADC , bla VIM and adeB were 64, 95, 61, 64 and 86 %, respectively. ISAba1 was found to be inserted into the 59 end of OXA-23 in 35 isolates (97 %). Of the AMEs, aadA1 (89 %) was the most prevalent, followed by aphA1 (75 %). The band patterns reproduced by REP-PCR showed that 34 out of 36 isolates belonged to one clone and two singletons were identified. The results confirmed that refractory A. baumannii isolates were widely distributed and warned the hospital infection control team to exert strict measures to control the infection. An urgent surveillance system should be implemented.

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“…Detection of bla OXA , bla PER , bla VEB , and bla GES genes ESBL-producing isolates were subjected to polymerase chain reaction (PCR) for detection of bla OXA , bla PER-1 bla VEB-1 , and bla GES-1 genes using the specific primers listed in Table 1 [18][19][20][21][22][23][24] . Total deoxyribonucleic acid (DNA) was extracted from bacterial isolates using an extraction kit (Bioneer, Daejeon, Korea).…”

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confidence: 99%

Distribution of the bla OXA , bla VEB-1 , and bla GES-1 genes and resistance patterns of ESBL-producing Pseudomonas aeruginosa isolated from hospitals in Tehran and Qazvin, Iran

Amirkamali

Naserpour-Farivar

Azarhoosh

et al. 2017

Rev. Soc. Bras. Med. Trop.

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Introduction:Pseudomonas aeruginosa is one of the most common nosocomial pathogens. The emergence of extended spectrum β-lactamases (ESBLs) has been increasingly reported as a major clinical concern worldwide. The main aim of the present study was to determine the distribution of bla OXA , bla PER-1, bla VEB-1, and bla GES-1 genes among ESBL-producing P. aeruginosa isolated from two distinct provinces in Iran. Methods: In this study, a total of 75 (27.5%) ESBL-producing isolates were identified from 273 P. aeruginosa isolates collected from patients in Qazvin and Tehran. Phenotypic detection of ESBLs and antimicrobial susceptibility testing were performed according to the Clinical and Laboratory Standards Institute guidelines. PCR and sequencing were employed to detect bla OXA-1 , bla OXA , bla GES-1, bla PER-1 , and bla VEB-1 genes. Isolate genetic relationships were evaluated by repetitive extragenic palindromic sequence-based PCR (REP-PCR). Results: In total, 59 (78.7%) of the ESBLproducing isolates showed multidrug resistance. The highest rates of susceptibility were observed against colistin (75 isolates, 100%) and polymyxin B (75, 100%) followed by amikacin (44, 58.7%), and piperacillin-tazobactam (40, 53.3%). The bla OXA-1 (37.3%) gene was the most common of the genes investigated, followed by bla OXA-4 (32%), bla GES-1 (16%), and bla VEB-1 (13.3%). REP-PCR identified three different genotypes: types A (89.3%), B (6.7%), and C (4%). Conclusions: We found a significant presence of bla OXA-1 , bla OXA-4 , bla GES-1, and bla VEB-1 genes among P. aeruginosa isolates, highlighting the need for suitable infection control strategies to effectively treat patients and prevent the further distribution of these resistant organisms.

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PCR-based DNA fingerprinting (REP-PCR, AP-PCR) and pulsed-field gel electrophoresis characterization of a nosocomial outbreak caused by imipenem- and meropenem-resistant Acinetobacter baumannii

Cited by 121 publications

References 35 publications

Class 1 integrons in non-clonal multidrug-resistant Acinetobacter baumannii from Iran, description of the new bla IMP-55 allele in In1243

Class 1 integrons in non-clonal multidrug-resistant Acinetobacter baumannii from Iran, description of the new bla IMP-55 allele in In1243

Limited genetic diversity and extensive antimicrobial resistance in clinical isolates of Acinetobacter baumannii in north-east Iran

Distribution of the bla OXA , bla VEB-1 , and bla GES-1 genes and resistance patterns of ESBL-producing Pseudomonas aeruginosa isolated from hospitals in Tehran and Qazvin, Iran

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