Boronic acid disk diffusion for the phenotypic detection of polymerase chain reaction-confirmed, carbapenem-resistant, gram-negative bacilli isolates

Elsherif, Rasha; Ismail, Dalia Kadry; Elawady, Sanaa; Jastaniah, Saddig D.; Almasaudi, Saad B.; Harakeh, Steve; Karrouf, Gamal

doi:10.1186/s12866-016-0754-z

Cited by 12 publications

(10 citation statements)

References 27 publications

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“…The most relevant epidemiologically and clinically important MβL types are VIM (Verona integrin-encoded MβL), IMP (imipenemase), NDM (New Delhi MβL), and SPM (Sao Paulo MβL). [56]…”

Section: Introductionmentioning

confidence: 99%

Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain

Joji¹,

Al-Rashed²,

Saeed

et al. 2019

J Lab Physicians

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INTRODUCTION: Carbapenem-resistant Pseudomonas aeruginosa has emerged as a life-threatening infectious agent worldwide. Carbapenemase genes are reported to be some of the most common mechanisms for carbapenem resistance in P. aeruginosa. No reports are available from the Kingdom of Bahrain about carbapenem resistance and the underlying cause. In this study, we determined to study the presence of the metallo-beta-lactamase (M β L) genes of VIM family and NDM-1 in carbapenem-resistant P. aeruginosa strains. METHODOLOGY: Fifty carbapenem-resistant P. aeruginosa isolates were obtained from three main hospitals of Bahrain. They were subjected to antimicrobial susceptibility testing by disc diffusion test. Subsequently, MβL was detected by imipenem-ethylene diamine tetraacetic acid (EDTA) combined disc test and conventional polymerase chain reaction. RESULTS: Among 50 P. aeruginosa strains, 40 (80%) were imipenem resistant. Among the 40 imipenem-resistant strains, 35 (87.5%) strains were positive for the imipenem-EDTA combined disc test, and 21 (52%) were carrying MβL genes. Nineteen (47.5%) strains were positive for the VIM gene; one (2.5%) strain was carrying the NDM-1 gene, while one strain was carrying both the VIM and NDM-1 genes. None of the imipenem sensitive strains carried the VIM or NDM-1 gene. CONCLUSION: This is the first study to report the presence of the VIM family gene and NDM-1 genes in imipenem-resistant P. aeruginosa isolates in the Kingdom of Bahrain. The study also confirms the multiple drug resistance by the MβL strains, attention should therefore from now on, be focused on prevention of further spread of such isolates by firm infection control measures, and to reduce its threat to public health.

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“…The most relevant epidemiologically and clinically important MβL types are VIM (Verona integrin-encoded MβL), IMP (imipenemase), NDM (New Delhi MβL), and SPM (Sao Paulo MβL). [56]…”

Section: Introductionmentioning

confidence: 99%

Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain

Joji¹,

Al-Rashed²,

Saeed

et al. 2019

J Lab Physicians

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“…Metallo-ß-lactamases such as imipemenase (IMP), Verona integrin-encoded-metallo-ß-lactamase (VIM) and New Delhi metallo-ß-lactamase (NDM) have been reported in Europe but currently appear to play a minor role (Evans et al 2013). The production of K. pneumoniae carbapenemase (KPC) and OXA-48 oxacillinase has been reported from the Middle East and South America, respectively (Elsherif et al 2016;Martinez et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Acinetobacter baumannii – a neglected pathogen in veterinary and environmental health in Germany

2018

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The emergence and global spread of drug resistant Acinetobacter (A.) baumannii is a cause of great concern. The current knowledge on antibiotic resistance in A. baumannii from animal origin is mostly based on few internationally published case reports, investigations of strain collections and several whole genome analyses. This lack of data results in a somewhat sketchy picture on how to assess the possible impact of drug resistant A. baumannii strains on veterinary and public health in Germany. Consequently, there is an urgent need to intensify the surveillance of A. baumannii in pet animals, the farm animal population and wildlife.

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“…Using sterile tips, 20 μl of 150 μg/mL PBA was dispensed onto one disc. 16 The second concentration was 600 μg/mL PBA, done as described by Elsherif, et al, 2016. Two ceftazidime (30 μg) discs were placed on the inoculated surface of the MHA plate at a distance of 25 mm, center to center.…”

Section: Phenotyping Of Ampc Using Disc Approximation Testmentioning

confidence: 99%

“…Using sterile tips, 20 μl of 600 μg/mL phenyl PBA was dispensed onto one disc. 17 After overnight incubation, the zone diameter around the antibiotic disc with added PBA and the antibiotic-containing disc alone were compared. An organism that demonstrates a defined increase (≥5-mm) in zone diameter around the antibiotic disc with added boronic acid was considered an AmpC producer.…”

Section: Phenotyping Of Ampc Using Disc Approximation Testmentioning

confidence: 99%

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Genetic detection of AmpC beta-lactamase among gram negative isolates “A Single Center Experience’’

Aal¹,

.²,

Rashed³

et al. 2021

EJI

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The misuse of β-Lactam antibiotics results in major problem, microbial resistance against these antibiotics by expression of β-lactamases, facing its use. AmpCs are one of the β-lactamases which confer resistance to penicillins, cephalosporins, cephamycins, and aztreonam, and are not affected by classic β-lactamase inhibitors. Plasmid-mediated AmpC β-lactamases pose a major challenge to infection control because the AmpC gene can be expressed in larger quantities and has a high transmissibility to other bacterial species. This study aimed to detect plasmid mediated AmpC β-lactamases in gram negative isolates in Assiut university hospital. It was performed on 120 cefoxtin resistant isolates obtained from 300-gram negative isolates using the disc diffusion method as a screening test for AmpC production. Since the presence of pAmpC is often associated with the presence of ESBLs, phenotypic detection of ESBL was done using combined disc method and vitek2 compact 15. Phenotypic detection of AmpC was done by disc approximation method and inhibitor-based method using phenyl boronic acid (PBA). Genotypic detection of 5 plasmid mediated AmpC genes families (MOX, CIT, DHA, EBC, and FOX) was done by multiplex PCR. Our result showed that Klebsiella pneumoniae (62.5%) and Escherichia coli (25.8%) were the most frequent isolates. Only 15.8 %, 12.5%, 17.5% resistant isolates to cefoxitin were positive by using disc approximation test, inhibitor-based method using PBA (150 g/mL), and PBA (600 g/mL), respectively. Out of the 120 Cefoxitin-resistant isolates, 22 isolates (18.3 %) were positive by multiplex PCR. CIT and MOX were solely detected in 45.5% and 4.5%, respectively. CIT and FOX together were detected in 45.5%, CIT and DHA together in 4.5%. No isolate was positive for EBC gene. Finally, boronic acid test using 600 μg/mL PBA with 30µg ceftazidime, as phenotypic method for detecting AmpC β- lactamases, was ranked very good for marking negative tests.

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Boronic acid disk diffusion for the phenotypic detection of polymerase chain reaction-confirmed, carbapenem-resistant, gram-negative bacilli isolates

Cited by 12 publications

References 27 publications

Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain

Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain

Acinetobacter baumannii – a neglected pathogen in veterinary and environmental health in Germany

Genetic detection of AmpC beta-lactamase among gram negative isolates “A Single Center Experience’’

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