MALDI-TOF MS detection of carbapenemase activity in clinical isolates of Enterobacteriaceae spp., Pseudomonas aeruginosa, and Acinetobacter baumannii compared against the Carba-NP assay

Chong, Patrick; McCorrister, Stuart; Unger, Mark; Boyd, David A.; Mulvey, Michael R.; Westmacott, Garrett

doi:10.1016/j.mimet.2015.01.024

Cited by 40 publications

(24 citation statements)

References 19 publications

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“…The biochemical Carba NP test yielded poor detection of OXA-48-type enzymes (sensitivity of around 80%), in contrast to the proposed method, which shows 100% sensitivity (9,17,18). The vast majority of articles reporting other MALDI-TOF MS assays focus on detecting carbapenemases (6,7,9), and few focus on detecting metallo-␤-lactamases (19) or oxacillinases (20).…”

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confidence: 90%

Rapid Detection of OXA-48-Producing Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry

et al. 2016

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A rapid and sensitive (100%) matrix-assisted laser desorption ionization؊time of flight mass spectrometry (MALDI-TOF MS) assay was developed to detect OXA-48-type producers, using 161 previously characterized clinical isolates. Ertapenem was monitored to detect carbapenem resistance, and temocillin was included in the assay as a marker for OXA-48-producers. Structural analysis of temocillin is described. Data are obtained within 60 min. Carbapenemase-producing Enterobacteriaceae (CPE) are increasingly being isolated worldwide. As these bacteria are difficult to detect and control, their spread is considered a public health concern (1, 2). Dissemination of the group D carbapenemases, particularly, bla OXA-48 isolates, is of major concern (3-5). These isolates are characterized by low carbapenem MICs and often go undetected by most phenotypic methods.The development of mass spectrometry techniques, such as matrix-assisted laser desorption ionizationϪtime of flight mass spectrometry (MALDI-TOF MS), has yielded sensitivity with faster turnaround times (6-10). The aim of this study was to determine whether a MALDI-TOF MS ertapenem assay can be used to identify and classify CPE producers, with particular emphasis on OXA-48-type isolates. We propose the use of temocillin as a resistance marker for OXA-48-type enzymes (11), as isolates harboring these types of carbapenemases show very high levels of resistance to temocillin (12, 13), and there is no specific inhibitor for group D carbapenemases. Phenylboronic acid (PBA) and dipicolinic acid (DPA) are used as inhibitors of, respectively, group A and B carbapenemases.A total of 161 characterized nonrepeat Enterobacteriaceae isolates, 117 of which carried a carbapenemase enzyme, were tested (Table 1). Seventy-two of the 161 isolates were genotypically characterized by the Antibiotic Resistance Surveillance Program of the Spanish National Center of Microbiology (14) All isolates were analyzed by the disk diffusion method (KPC/ MBL and OXA-48 confirm kit; Rosco Diagnostica A/S, Taastrup, Denmark) with disks containing meropenem (10 g), meropenem plus PBA, meropenem plus DPA, meropenem plus cloxacillin (CL), and temocillin (30 g). A 0.5-McFarland suspension of the isolates on Mueller-Hinton II agar plates (Becton Dickinson, USA) was used, followed by an overnight incubation of the plates at 37°C. The susceptibility cutoff used for temocillin was Ͻ12 mm (12).For the MALDI-TOF MS assay, isolates were previously grown overnight at 37°C in Trypticase soy agar with 5% sheep blood (TSA II) (Becton Dickinson). The hydrolysis of ertapenem (0.5 mg/ml Invaz [Merck & Co., Inc.], 0.001% SDS, 50 mM NH 4 HCO 3 [pH 7.0]) (6, 9) and temocillin (1.5 mg/ml Negaban [Eumedica], 0,001% SDS, 50 mM NH 4 HCO 3 [pH 7.0]) (9) was evaluated after resuspension of the bacteria that filled a 1-l inoculation loop in 10 l of the antibiotic solution in an Eppendorf tube and incubation of the suspension at 37°C under agitation. When inhibitors were used, bacteria were resuspended in 5 l of ertapenem (1 mg/ml ...

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Rapid Detection of OXA-48-Producing Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry

et al. 2016

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“…The Carba NP test has been commercialized through the Rapidec Carba NP system (bioMérieux, France) (25,26), facilitating the procedure and hands-on time. However, false-positive and -negative results have been detected for OXA-48-like variants, in a more significant number than with the in-house Carba NP test (27). To date, the different variations of the Carba NP test has been used to detect not only ␤-lactamase resistance but also aminoglycoside and polymyxin resistance, all mediated by enzymes (28,29).…”

Section: Introductionmentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

2018

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SUMMARY Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied in recent years for first-line identification of pathogens in clinical microbiology because it is simple to use, rapid, and accurate and has economic benefits in hospital management. The range of clinical applications of MALDI-TOF MS for bacterial isolates is increasing constantly, from species identification to the two most promising applications in the near future: detection of antimicrobial resistance and strain typing for epidemiological studies. The aim of this review is to outline the contribution of previous MALDI-TOF MS studies in relation to detection of antimicrobial resistance and to discuss potential future challenges in this field. Three main approaches are ready (or almost ready) for clinical use, including the detection of antibiotic modifications due to the enzymatic activity of bacteria, the detection of antimicrobial resistance by analysis of the peak patterns of bacteria or mass peak profiles, and the detection of resistance by semiquantification of bacterial growth in the presence of a given antibiotic. This review provides an expert guide for MALDI-TOF MS users to new approaches in the field of antimicrobial resistance detection, especially possible applications as a routine diagnostic tool in microbiology laboratories.

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“…Bu çalışmalarda testlerin duyarlılıkları % 77-100, özgüllükleri % 94-100 arasında değişmektedir. Yalancı negatif sonuçlar çoğunlukla OXA-48 pozitif kökenlerde tespit edilmiştir (3,7,14,12,13,16,18,21,24) . Çalışmamızda ise KPC, NDM ve IMP tipi karbapenemazların VITEK MS ile tespiti 2 saat gibi kısa bir sürede sağlanmıştır.…”

Section: Discussionunclassified

“…Son yıllarda araştırıcılar bu sistemlerin sadece mikroorganizmaların tanımlanmasında değil aynı zamanda antibiyotik duyarlılık testlerinde de kullanılabileceğini gösteren çalışmalar bildirmektedir. Özellikle beta laktamaz ve karbapenemazların antibiyotikleri hidroliz etmesi sonucu oluşan değişikliklerin MALDI-TOF MS aracığıyla saptanmasına yönelik çalışmalar tüm dünyada hız kazanmıştır (3,7,(12)(13)(14)16,18,21,24) . Çalışmamızda VITEK-MS (bioMérieux SA, Marcyl'E´toile, Fransa) sistemi kullanılarak ertapenem hidrolizi aracığı ile karbapenemaz üretiminin hızlı saptanması amaçlanmıştır.…”

Section: Introductionunclassified

Rapid Detection of Carbapenemase Detection by VITEK-MS

Gelmez¹,

Can²,

Hasdemir³

et al. 2019

Ankem Derg

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Son yıllarda karbapenemaz üreten Enterobacteriaceae kaynaklı enfeksiyonların dünya çapında artması önemli halk sağlığı sorunudur. Araştırıcılar, mikroorganizmanın kısa sürede tanımlanmasına olanak sağlayan MALDI-TOF sisteminin antibiyotik duyarlılık testlerinde de kullanılabileceğini gösteren çalışmalar bildirilmektedir. Çalışmamızda karbapenemaz üretiminin hızlı saptanmasına yönelik olarak ertapenemin karbapenemazlar tarafından hidrolizinin VITEK-MS (bioMérieux, Fransa) sistemi tarafından tespit edilebilirliği değerlendirilmiştir. Çalışmamıza çeşitli klinik örneklerden izole edilen karbapenem dirençli (n=86) ve duyarlı (n=20) toplam 106 Klebsiella pneumoniae kökeni dahil edilmiştir. KPC, NDM, IMP, VIM ve OXA-48 genlerinin varlığını PCR ile tespit edilmiştir. Bakteri süspansiyonları 4 McFarland bulanıklığında hazırlanmış, 0.5 µg/ml'lik ertapenem ile muamele edildikten sonra iki ve dört saat inkübe edilip analiz edilmek üzere VITEK-MS RUO (bioMérieux, Fransa) sistemine aktarılmıştır. Karbapenem dirençli 86 K.pneumoniae kökeninin bla OXA-48 (n=39), bla NDM (n= 37), bla IMP (n=8) ve bla KPC (n=2) karbapenemaz genlerini taşıdığı tespit edilmiştir. Tüm antibiyotiklere duyarlı olan kökenlerde hiçbir karbapenemaz genine rastlanmamıştır. Ertapenemin tek başına sahip olduğu spektrumlar çalışma kökenlerinin ertapenem ile muamele edilip inkübe edilmesi sonucu elde edilen spektrumlar ile karşılaştırıldığında; iki saatlik inkübasyon sonrasında NDM, IMP ve KPC geni pozitif kökenlerin tamamında ertapenem hidrolizi gözlenirken, OXA-48 geni pozitif olan 14 kökende ertapenem hidrolizinin zayıf olduğu tespit edilmiştir. Dört saatlik inkübasyon sonrası OXA-48 geni pozitif olan kökenlerin sadece % 64.1'inde ertapenem hidrolizi gözlemlenmiştir. İki ve dört saat inkübasyon sonrası karbapenemaz geni içermeyen kökenlerin hiçbirinde ertapenem hidrolizi gözlemlenmemiştir. Rutin laboratuvarda mikroorganizmaların hızlı tespitinde önemli başarı sağlayan MALDI-TOF sistemlerinin antibiyotik duyarlılık testlerinde de benzer başarı göstermesi oldukça ümit vericidir. Ancak OXA-48 pozitif kökenlerin saptanmasını kolaylaştıracak farklı protokollerin geliştirilmesi gereklidir. MALDI-TOF sistemleri ile kısa sürede, düşük maliyetle karbapenemaz üretiminin tespit edilmesi özellikle salgınların çok daha kısa sürede kontrol altına alınmasında önemli katkı sağlayacaktır.

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MALDI-TOF MS detection of carbapenemase activity in clinical isolates of Enterobacteriaceae spp., Pseudomonas aeruginosa, and Acinetobacter baumannii compared against the Carba-NP assay

Cited by 40 publications

References 19 publications

Rapid Detection of OXA-48-Producing Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry

Rapid Detection of OXA-48-Producing Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

Rapid Detection of Carbapenemase Detection by VITEK-MS

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