Comparative Evaluation of Multiplex P CR and Routine Laboratory Phenotypic Methods for Detection of Carbapenemases among Gram Negative Bacilli

Solanki, Rachana; Vanjari, Lavanya; Subramanian, S; Lakshmi, .

doi:10.7860/jcdr/2014/10794.5322

Cited by 29 publications

(23 citation statements)

References 18 publications

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“…7%) , similar to finding by Vijayakumar et al (2016) from Tamil Nadu and Solanki R et al (2014) from Hyderabad i.e. 19.2% and 36.4% respectively(34,35). Some of the studies in India e.g.…”

supporting

confidence: 85%

Molecular Epidemiology of Metallo Beta Lactamases in Acinetobacter Baumannii at a Tertiary Care Hospital

Mohammed¹,

Singh²

2019

IJMBS

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Background: Carbapenem resistance mediated by metallo beta lactamases (MBL) in Acinetobacter baumannii is a global challenge due to its rapid spread and limited therapeutic options. Objective: To determine the prevalence of MBL in A. baumannii isolates in hospitalized patients by both phenotypic and genotypic methods. Materials and Methods: The clinical samples were collected from inpatients and subcultured on routine culture media for growth. Identification of bacteria along with antimicrobial sensitivity testing was done by VITEK -2 Compact (bioMerieux). Antibiotics that were not tested by VITEK-2 were tested manually by Kirby-Bauer disk diffusion method according to CLSI 2017 and EUCAST 2016 guidelines. The isolates which were resistant to carbapenem (imipenem and/ or meropenem) were tested by phenotypic (imipenem-EDTA combined disk method) and genotypic method for presence of common metallo beta lactamases genes (blaIMP, blaNDM, blaGIM, blaVIM, blaSPM and blaSIM). Results: 84 non duplicate A.baumannii were isolated out of 947 pathogenic gram negative isolates. Majority (47.6%) of isolates were obtained from tracheostomy/endotracheal/bronchoalveolar lavage (TT/ET/BAL) followed by sputum (21.4%). None of the isolates were found to be resistant to colistin and tigecycline. 73 (86.9%) isolates were found to be carbapenem resistant, among these 60 (82.2%) were found to be MBL positive by phenotypic and 32 (43.2%) by genotypic method. MBL genes detected were blaNDM (39.7%), blaGIM (2.7%) and blaVIM (1.4%). None of the isolates were positive for blaIMP, blaSPM and blaSIM. Conclusion: The prevalence of MBL in carbapenem resistant isolates of A.baumannii was 87.7%. blaNDM was the most common gene detected. No significant difference was found in the ability of phenotypic and genotypic methods for MBL detection. The resistance rate of the A.baumannii is high for most antibiotics except for polymyxins (E&B) and tigecycline. Key words: Metallo beta Lactamases, Acinetobacter baumannii, Carbapenem.

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“…7%) , similar to finding by Vijayakumar et al (2016) from Tamil Nadu and Solanki R et al (2014) from Hyderabad i.e. 19.2% and 36.4% respectively(34,35). Some of the studies in India e.g.…”

supporting

confidence: 85%

Molecular Epidemiology of Metallo Beta Lactamases in Acinetobacter Baumannii at a Tertiary Care Hospital

Mohammed¹,

Singh²

2019

IJMBS

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“…But they cannot hydrolyze aztreonam. They can be inhibited by EDTA [ 1 , 10 , 12 ]. E-test MBL strips use the principle of inhibition of metallo-β-lactamases by EDTA when IPM is used as a substrate.…”

Section: Discussionmentioning

confidence: 99%

“…Automated antibiotic susceptibility systems were found to be unreliable for detection of carbapenem resistance. These automated systems either over or under reported carbapenem resistance [ 2 , 12 ]. More practical molecular methods are needed.…”

Section: Discussionmentioning

confidence: 99%

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Phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae in a tertiary-level reference hospital in Turkey

Baran

Aksu

2016

Ann Clin Microbiol Antimicrob

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BackgroundEnterobacteriaceae are among the most common pathogens that are responsible for serious community-acquired, hospital-acquired, and health-care associated infections. The emergence and spread of carbapenem-resistant Enterobacteriaceae (CRE) have become an increasing concern for healthcare services worldwide. Infections caused by these bacteria have been associated with significant morbidity and mortality and treatment options have been limited. The rapid and accurate detection of carbapenem resistance in these bacteria is important for infection control. The aim of this study was to investigate the phenotypic and genotypic features of CRE strains isolated in a tertiary-level reference hospital in Turkey.MethodsA total of 181 CRE strains were included in the study. Antimicrobial susceptibility rates were tested using Vitek 2 system. Modified Hodge test (MHT) was performed using meropenem and ertapenem discs. Metallo-β-lactamase antimicrobial gradient test (E-test MBL strips) were used for evaluation of metallo-β-lactamase production. A multiplex PCR was used for detection of carbapenems resistance genes (IMP, VIM, KPC, NDM-1 and OXA-48).ResultsThe OXA-48 gene was detected in 86 strains, NDM-1 gene in six strains, VIM gene in one strain. IMP and KPC genes were not identified. Three strains produced both OXA-48 and NDM-1 and one strain produced both OXA-48 and VIM. In two patients more than one genus of OXA-48 positive CREs was isolated. Ninety-two of the isolates were multidrug-resistant. One hundred and ten isolates were MHT with meropenem (MEM-MHT) positive and 109 isolates were MHT with ertapenem (ERT-MHT) positive. Nine of the isolates were positive with E-test MBL strips. The sensitivity of MEM-MHT and ERT-MHT for detection of OXA-48 was 70.9 and 70.6 %, respectively. MEM-MHT was found highly discriminative for OXA-48 Escherichia coli (p < 0.001). The sensitivity of E-test MBL for NDM-1 was 66.7 %. A statistically significant correlation was observed between OXA-48 gene and MHT positivity and between NDM-1, VIM gene and E-test MBL positivity (p < 0.001).ConclusionsOXA-48 gene is spreading rapidly to many different species of Enterobacteriaceae in the hospital environment. While OXA-48 is still the most common source of carbapenem resistance in Enterobacteriaceae in our country, NDM-1 is increasingly being isolated from patients without a history of foreign contact.

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“…The products must be of different sizes and can be visualized either by gel electrophoresis, if from conventional PCR, or by addition of different dyes for RT-PCR. Multiplex PCRs are often designed to detect different genes, all relating to the same resistance phenotype such as detection of the most prevalent beta-lactamases present in Gram-negative bacteria which are involved in resistance to cephalosporins (15) or carbapenems (16,17). Therefore, by screening for these genes simultaneously, considerable time and effort can be saved in detecting the possible mechanism(s) responsible for the resistance phenotype.…”

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confidence: 99%

Molecular Methods for Detection of Antimicrobial Resistance

Anjum

Zankari

Hasman

2018

Antimicrobial Resistance in Bacteria From Livestock and Companion Animals

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Comparative Evaluation of Multiplex P CR and Routine Laboratory Phenotypic Methods for Detection of Carbapenemases among Gram Negative Bacilli

Cited by 29 publications

References 18 publications

Molecular Epidemiology of Metallo Beta Lactamases in Acinetobacter Baumannii at a Tertiary Care Hospital

Molecular Epidemiology of Metallo Beta Lactamases in Acinetobacter Baumannii at a Tertiary Care Hospital

Phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae in a tertiary-level reference hospital in Turkey

Molecular Methods for Detection of Antimicrobial Resistance

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