Dipicolinic acid-based disk methods for detection of metallo-β-lactamase–producing Pseudomonas spp. and Acinetobacter spp.

Shin, Kyeong Seob; Son, Bo Ra; Hong, Seung Bok; Kim, Jungmin

doi:10.1016/j.diagmicrobio.2008.04.015

Cited by 24 publications

(17 citation statements)

References 9 publications

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“…In a previous study, it was reported that the best results were observed when the DPA content in the disk was 835 g for DDPTs and DPTs (17). In our study, use of a disk with 835 g of DPA did not improve the sensitivity of the MEM-DPA DDPT test (Table 1, method A versus method C), whereas a 250-g DPA disk achieved high sensitivity (97.7%) when used with an IPM disk (method B).…”

Section: Discussionmentioning

confidence: 99%

“…Regarding the DPA content, the commercial tablet contains 250 g, whereas Shin et al (17) used an 835-g disk. Recently, it was reported that a meropenem (MEM)-DPA (1,000 g) DPT using commercial tablets (Rosco Diagnostica) showed 100% sensitivity and specificity for the detection of Enterobacteriaceae isolates with VIM-1-and IMP-type MBL (4).…”

mentioning

confidence: 99%

“…and VIM-2-and IMP-1-producing Acinetobacter spp. (17), but the study did not include SIM-1-producing Acinetobacter spp. isolates, which are present in Korea (13) and China (21).…”

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confidence: 99%

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Evaluation of Double-Disk Potentiation and Disk Potentiation Tests Using Dipicolinic Acid for Detection of Metallo-β-Lactamase-Producing Pseudomonas spp. and Acinetobacter spp

Yong

Lee

Jeong³

et al. 2012

J Clin Microbiol

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bAccurate detection of metallo-␤-lactamase (MBL)-producing Pseudomonas spp. and Acinetobacter spp. became very important with the increasing prevalence of carbapenem-nonsusceptible clinical isolates. The performance of phenotypic MBL detection methods may depend on the types of MBL and the characteristics of the isolates. A high false-positive rate is a problem with EDTA-based MBL detection methods. We evaluated the performance of double-disk potentiation tests (DDPTs) and disk potentiation tests (DPTs) with dipicolinic acid (DPA) using 44 isolates of Pseudomonas spp. and Acinetobacter spp. producing IMP-1-like, VIM-2-like, and SIM-1 type MBLs. Also, we characterized P. aeruginosa isolates with positive imipenem (IPM)-DPA DDPT, but negative meropenem (MEM)-DPA DDPT, and determined possibility of improving a DDPT by using MacConkey agar. Among five different DDPT methods, the IPM-DPA 250-g method showed the highest sensitivity (97.7%) and specificity (100%). Among four DPT tests, the highest sensitivity (100%) was shown by the IPM-EDTA 1,900-g disk method, but the specificity was very low (11.4%). Five of six P. aeruginosa isolates with false-negative DDPTs with MEM-DPA 250-g disks carried bla IMP-6, and the high level resistance to MEM (MIC > 512 g/ml) was reduced by the presence of phenylalanine arginine ␤-naphtylamide. Improvement of DDPTs was observed when MacConkey agar was used instead of Mueller-Hinton agar. In conclusion, DPA is a better MBL inhibitor than EDTA for detection of Pseudomonas spp. and Acinetobacter spp. with IMP-1-like, VIM-2-like, and SIM-1-type MBLs. In DPA DDPTs, IPM disks perform better than MEM disks when the isolates are highly resistant to MEM due to the overexpression of efflux pumps. C arbapenems are often used as "antibiotics of last resort" when patients with infections become gravely ill or are suspected of harboring resistant bacteria (14). However, carbapenem-resistant Gram-negative bacilli have been increasingly reported worldwide. In Korea, carbapenem-resistant isolates of Enterobacteriaceae remains very rare, but during the period of 2001 to 2009, imipenem (IPM)-resistant Pseudomonas aeruginosa and Acinetobacter spp. increased from 17 to 26% and from 5 to 51%, respectively (10). A significant proportion of the resistance in P. aeruginosa is due to the production of VIM-2-like and IMP-1-like metallo-␤-lactamase (MBL), whereas in Acinetobacter spp. the majority are due to the production of OXA-type carbapenemase, although VIM-2-like, IMP-1-like, and SIM-1 enzyme-producing isolates have also been detected (8).Various inhibitor-based methods are commonly used to detect MBLs, but these techniques are not highly sensitive or highly specific. A disk potentiation test (DPT) is also called a combined disk test. A DPT using imipenem (IPM) and EDTA (IPM-EDTA DPT) showed a high false-positive rate with Acinetobacter spp. due to growth inhibition by EDTA alone (20). A double-disk potentiation test (DDPT) is also known as a double-disk synergy test. To reduce false-positive results from t...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Evaluation of Double-Disk Potentiation and Disk Potentiation Tests Using Dipicolinic Acid for Detection of Metallo-β-Lactamase-Producing Pseudomonas spp. and Acinetobacter spp

Yong

Lee

Jeong³

et al. 2012

J Clin Microbiol

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“…The library was screened for bla BlaB in LB-chloramphenicol medium containing 50 g/ml ampicillin. Isolated clones were tested for production of carbapenemase activity on plates by facing disks containing imipenem and dipicolinic acid (DPA) as described below (39). DNA constructions were verified by sequencing at the DNA Sequencing Service of the University of Maine (Orono, ME).…”

Section: Methodsmentioning

confidence: 99%

Carbapenem Resistance in Elizabethkingia meningoseptica Is Mediated by Metallo-β-Lactamase BlaB

González

Vila

2012

Antimicrob Agents Chemother

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Elizabethkingia meningoseptica, a Gram-negative rod widely distributed in the environment, is resistant to most ␤-lactam antibiotics. Three bla genes have been identified in E. meningoseptica, coding for the extended-spectrum serine-␤-lactamase CME (class D) and two unrelated wide-spectrum metallo-␤-lactamases, BlaB (subclass B1) and GOB (subclass B3). E. meningoseptica is singular in being the only reported microorganism possessing two chromosomally encoded MBL genes. Real-time PCR and biochemical analysis demonstrate that the three bla genes are actively expressed in vivo as functional ␤-lactamases. However, while CME elicits cephalosporin resistance, BlaB is the only ␤-lactamase responsible for E. meningoseptica resistance to imipenem, as GOB activity is masked by higher cellular levels of BlaB. On the other hand, we demonstrate that bla BlaB expression is higher in the stationary phase or under conditions that mimic the nutrient-limiting cerebrospinal fluid colonized by E. meningoseptica in human meningitis.

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“…A modified Hodge test [18] and MBL disk test [19] were performed for carbapenemase and MBL screening, respectively. …”

Section: Bacterial Strains Identification Antimicrobial Susceptibilimentioning

confidence: 99%

Characterization ofAcinetobacter baumanniiCo-producing Carbapenemases OXA-23 and OXA-66, andarmA16S Ribosomal RNA Methylase at a University Hospital in South Korea

Jeong

Son

Shin

et al. 2011

Korean J Clin Microbiol

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Background:In the present study, the resistance mechanisms against carbapenems and aminoglycosides for 23 strains of multi-drug-resistant Acinetobacter baumannii isolated at a university hospital were investigated. Methods: The minimal inhibitory concentrations (MICs) were determined via broth microdilution or Etest. The genes encoding OXA-type carbapenemases and 16S rRNA methylase were identified using multiplex PCR, and the amplified products were sequenced. Conjugation experiments were conducted, and an epidemiologic study was performed using enterobacterial repetitive intergenic consensus (ERIC)-PCR. Results: In the isolates, the MICs of the tested aminoglycosides, including arbekacin, were ＞1024 μg/ mL; the MICs of aztreonam, cefepime, ceftazidime, and ciprofloxacin ranged from 64 to 128 μg/mL; and the MICs of carbapenem ranged from 32 to 64 μg/ mL, as determined through the broth microdilution test. According to the E-test, the MICs of ampicillin/ sulbactam and colistin were 8 and 0.25 to 0.38 μg/ mL, respectively. Sequence analysis confirmed that all of the isolates expressed carbapenemases OXA-23 and OXA-66, as well as armA 16S rRNA methylase. In addition, ISAba1 was identified upstream of the gene encoding OXA-23. OXA-23 and armA were not transferred to Escherichia coli J53 cells in the transconjugation experiments. ERIC-PCR molecular fingerprinting produced a single pattern in all cases. Conclusion:The co-production of OXA-23 and armA 16S rRNA methylase may be attributed to the multidrug resistance of the A. baumannii isolates in the present study. Stricter surveillance and more rapid detection are necessary to prevent the spread of this type of resistance in the future. (Korean J Clin Microbiol 2011;14:67-73)

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Dipicolinic acid-based disk methods for detection of metallo-β-lactamase–producing Pseudomonas spp. and Acinetobacter spp.

Cited by 24 publications

References 9 publications

Evaluation of Double-Disk Potentiation and Disk Potentiation Tests Using Dipicolinic Acid for Detection of Metallo-β-Lactamase-Producing Pseudomonas spp. and Acinetobacter spp

Evaluation of Double-Disk Potentiation and Disk Potentiation Tests Using Dipicolinic Acid for Detection of Metallo-β-Lactamase-Producing Pseudomonas spp. and Acinetobacter spp

Carbapenem Resistance in Elizabethkingia meningoseptica Is Mediated by Metallo-β-Lactamase BlaB

Characterization ofAcinetobacter baumanniiCo-producing Carbapenemases OXA-23 and OXA-66, andarmA16S Ribosomal RNA Methylase at a University Hospital in South Korea

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