A MALDI TOF MS-based minisequencing method for rapid detection of TEM-type extended-spectrum beta-lactamases in clinical strains of Enterobacteriaceae

Ikryannikova, Larisa N.; Shitikov, Egor; Zhivankova, D.G.; Ilina, Elena; Edelstein, Mikhail V.; Govorun, Vadim M.

doi:10.1016/j.mimet.2008.07.005

Cited by 27 publications

(21 citation statements)

References 15 publications

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“…To date, more than 400 different structures of β-lactamase have been discovered and divided into 4 classes according to the Bush method (Bush and Jacoby, 2010). The structural diversity of β-lactamases is a challenge to screening techniques used to directly identify the specific AA or nucleic acid sequences of the enzyme: PCR (López-Cerero et al, 2011;Voets et al, 2011), immunoassays (Hujer et al, 2002;Hujer et al, 2009), spectrophotometric assays (Gao et al, 2003;Perret, 1954;Xing et al, 2005), isoelectric focusing (Paterson et al, 2001), and mass spectrometry-based methods (Ikryannikova et al, 2008). On the other hand, indirect methods have been successfully developed that measure the activity of β-lactamase by monitoring the enzymatic reactions with its specific substrate.…”

Section: Introductionmentioning

confidence: 99%

A rapid HPLC method for indirect quantification of β-lactamase activity in milk

Zhou

Wang

Zhao

et al. 2015

Journal of Dairy Science

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To circumvent the strictly regulated limits of antibiotics in milk, illegal addition of β-lactamase to lower the antibiotic levels in milk has been reported recently in China. Herein, we describe a fast, sensitive, and robust HPLC-UV method for the determination of β-lactamase activity in milk, based on an indirect quantification strategy. The test milk sample was mixed with a known amount of penicillin G, a specific substrate of β-lactamase. After incubation, an aliquot of the mixture was injected into the HPLC-UV system to quantify the remaining penicillin G in less than 10 min. Comparative analysis of the amount of penicillin G before and after incubation was used to indirectly deduce the activity of β-lactamase in the test sample. This method was successfully applied to milk products with different fat percentages, resulting in a detection limit of 0.6 U/mL. Good recoveries, ranging from 94 to 105%, were obtained from blank milk samples spiked with β-lactamase at levels of 2 to 50 U/mL, with relative standard deviations <6%. A good correlation was demonstrated between the HPLC method and the conventional culture-based assay. Using this method, the activity changes in β-lactamase during milk pasteurization, sterilization, and storage were investigated. The advantages of low-cost, accurate quantification and easily accessible instrumentation make the proposed method an ideal alternative for high-throughput routine analysis in the dairy industry.

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Section: Introductionmentioning

confidence: 99%

A rapid HPLC method for indirect quantification of β-lactamase activity in milk

Zhou

Wang

Zhao

et al. 2015

Journal of Dairy Science

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“…Currently, multiple applications of MALDI-TOF MS are being pursued for the detection of resistance mechanisms within Ent . These applications include: identification of the antibiotic itself and its modified/degradated counterparts (Burckhardt & Zimmermann, 2011; Hooff, et al, 2012; Hrabak, et al, 2012; Hrabak, Walkova, Studentova, Chudackova, & Bergerova, 2011; Sparbier, Schubert, Weller, Boogen, & Kostrzewa, 2012), detection of the resistance proteins within the cell (Cai, Hu, Zhang, Zhou, & Chen, 2012; Camara & Hays, 2007; Schaumann, et al, 2012), and discovery of mutations within resistance genes through mini-sequencing (Ikryannikova, et al, 2008). …”

Section: Mass Spectrometry-based Methodologiesmentioning

confidence: 99%

“…focused on three nucleotide positions that correspond to the amino acid positions 104, 164, and 238, as mutations in these gene regions can confer an ESBL phenotype in TEM-type β-lactamases. The system identified polymorphisms accurately at these three sites (Ikryannikova, et al, 2008). In another investigation, Stürenburg et al .…”

Section: Mass Spectrometry-based Methodologiesmentioning

confidence: 99%

Non-phenotypic tests to detect and characterize antibiotic resistance mechanisms in Enterobacteriaceae

Lupo¹,

Papp-Wallace²,

Sendi³

et al. 2013

Diagnostic Microbiology and Infectious Disease

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In the past two decades, we have observed a rapid increase of infections due to multidrug-resistant Enterobacteriaceae. Regrettably, these isolates possess genes encoding for extended-spectrum β-lactamases (e.g., blaCTX-M, blaTEM, blaSHV) or plasmid-mediated AmpCs (e.g., blaCMY) that confer resistance to last-generation cephalosporins. Furthermore, other resistance traits against quinolones (e.g., mutations in gyrA and parC, qnr elements) and aminoglycosides (e.g., aminoglycosides modifying enzymes and 16S rRNA methylases) are also frequently co-associated. Even more concerning is the rapid increase of Enterobacteriaceae carrying genes conferring resistance to carbapenems (e.g., blaKPC, blaNDM). Therefore, the spread of these pathogens puts in peril our antibiotic options. Unfortunately, standard microbiological procedures require several days to isolate the responsible pathogen and to provide correct antimicrobial susceptibility test results. This delay impacts the rapid implementation of adequate antimicrobial treatment and infection control countermeasures. Thus, there is emerging interest in the early and more sensitive detection of resistance mechanisms. Modern non-phenotypic tests are promising in this respect, and hence, can influence both clinical outcome and healthcare costs. In this review, we present a summary of the most advanced methods (e.g., next-generation DNA sequencing, multiplex PCRs, real-time PCRs, microarrays, MALDITOF MS, and PCR/ESI MS) presently available for the rapid detection of antibiotic resistance genes in Enterobacteriaceae. Taking into account speed, manageability, accuracy, versatility, and costs, the possible settings of application (research, clinic, and epidemiology) of these methods and their superiority against standard phenotypic methods are discussed.

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“…128 Overall, the main advantages of MALDI-TOF MS are its rapidity, very low costs, and consistency. [129][130][131][132] Recently, this system has also been applied to (i) recognize antibiotic degradation owing to resistance proteins [133][134][135][136][137][138][139][140][141][142] ; (ii) discover mutations within resistance genes through DNA minisequencing 142 ; and (iii) study clonality of Gram-negative pathogens (eg, detection of ST131 or ST405 E coli). 143,144 Numerous studies have assessed specifically the usefulness of MALDI-TOF MS for the identification of b-lactam degradation products in the presence of hydrolyzing b-lactamases, [137][138][139][140][141]145,146 including directly from positive blood cultures.…”

Section: Matrix-assisted Laser Desorption Ionization-time Of Flight Mmentioning

confidence: 99%

The Changing Role of the Clinical Microbiology Laboratory in Defining Resistance in Gram-negatives

Endimiani

Jacobs

2016

Infectious Disease Clinics of North America

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A MALDI TOF MS-based minisequencing method for rapid detection of TEM-type extended-spectrum beta-lactamases in clinical strains of Enterobacteriaceae

Cited by 27 publications

References 15 publications

A rapid HPLC method for indirect quantification of β-lactamase activity in milk

A rapid HPLC method for indirect quantification of β-lactamase activity in milk

Non-phenotypic tests to detect and characterize antibiotic resistance mechanisms in Enterobacteriaceae

The Changing Role of the Clinical Microbiology Laboratory in Defining Resistance in Gram-negatives

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