β- Laktamazlar ve Genotipik Testler
Özet
Antimikrobiyal direnç genlerinin moleküler olarak tiplendirilmesi, antibiyotik duyarlılık testleri ile birlikte enfeksiyon hastalıkları ile mücadelede oldukça önemlidir. Rutin laboratuvarlarda fenotipik yöntemler öncelikli olarak tercih edilse de bu yöntemler genellikle zaman alıcı olup antibiyotik direncini saptamadaki yetenekleri sınırlıdır. Son yıllarda moleküler biyolojideki ilerlemeler patojenlerin ve direnç genlerinin saptanmasında kullanılabilecek birçok yöntemin geliştirilmesini sağlamıştır. Polimeraz zincir reaksiyonu, izotermal amplifikasyon, mikroarray ve yeni nesil dizileme yüksek duyarlılık ve özgüllüğe sahip testlerdir. Moleküler testler özellikle sendromik bir yaklaşımla kullanıldığında tanı, tedavi ve enfeksiyon kontrol önlemlerinin gerçekleştirilmesinde önemli yarar sağlar. Ayrıca moleküler testler, olası bir salgın durumunda dirençli bakterileri kontrol altına alma ve epidemiyolojik verilerin oluşmasında hızlı çözümler sunar. Antibiyotik direnç genlerinin moleküler tanımlanması yerel, ulusal ve küresel antibiyotik direnç sürveyansı için de kullanılmaktadır. Bu derlemede tüm dünyada ciddi boyutlara ulaşan ß-laktamaz üreten bakterilerin saptanmasında kullanılan moleküler yöntemler tartışılacaktır.
Molecular characterization of antimicrobial resistance (AMR) genes, along with antimicrobial susceptibility testing, is crucial in combating infectious diseases. While phenotypic methods are primarily preferred in routine laboratories, these methods are generally time-consuming and have limited ability to detect antibiotic resistance. Recent advances in molecular biology have led to the development of several methods for detecting pathogens and resistance genes. Techniques such as polymerase chain reaction (PZR), isothermal amplification, microarrays, and next-generation sequencing are highly sensitive and specific tests. Molecular tests, especially when used with a syndromic approach, provide significant benefits in diagnosis, treatment, and the implementation of infection control measures. Additionally, molecular tests offer rapid solutions for controlling resistant bacteria in the event of an outbreak and for generating epidemiological data. The molecular characterization of antimicrobial resistance genes is also used for local, national, and global antimicrobial resistance surveillance. This review will discuss molecular methods used in the detection of ß-lactamase-producing bacteria, which have become a serious issue worldwide.
Referanslar
Bush K, Jacoby GA. Updated functional classification of ß-lactamases. Antimicrob Agents Chemother. 2010;54(3):969-76.
Zhuang Q, Guo H, Peng T, et al. Advances in the detection of β-lactamase: A review. Int J Biol Macromol. 2023;251:126159.
Lawrence J, O'Hare D, van Batenburg-Sherwood J, Sutton M, Holmes A, Rawson TM. Innovative approaches in phenotypic ß-lactamase detection for personalised infection management. Nat Commun 2024; 15, 9070 https://doi.org/10.1038/s41467-024-53192-7
Elbehiry A, Marzouk E, Abalkhail A, et al. Detection of antimicrobial resistance via state-of-the-art technologies versus conventional methods. Front Microbiol. 2025;16:1549044.
Noster J, Thelen P, Hamprecht A. Detection of Multidrug-Resistant Enterobacterales-From ESBLs to Carbapenemases. Antibiotics (Basel). 2021 Sep 21;10(9):1140.
Priyanka Uprety, Thomas J Kirn. 2023. Molecular Detection of Antibacterial Drug Resistance In: Carroll KC, Pfaller MA Manual of Clinical Microbiology, 12th Edition. ASM Press, Washington, DC.(BU REF YAZIMI DOĞRU MU?)
Mullis K, Faloona F, Scharf S, Saiki R, Horn G, Erlich H. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb Symp Quant Biol. 1986;51(1):263-73.
Rahman M, Waseka AJ. Clinical Laboratory and Molecular Detection of Extended Spectrum ß lactamases: A Review Update. Bangladesh Journal of Infectious Diseases, 2015;1(1):12.
Sahoo R, Jadhav S, Nema V. Journey of technological advancements in the detection of antimicrobial resistance. J Formos Med Assoc. 2024 Apr;123(4):430-441.
Rood IGH, Li Q. Review: Molecular detection of extended spectrum-β-lactamase- and carbapenemase-producing Enterobacteriaceae in a clinical setting. Diagn Microbiol Infect Dis. 2017;89(3):245-250.
Zhuang Q, Guo H, Peng T, et al. Advances in the detection of β-lactamase: A review. Int J Biol Macromol. 2023;251:126159.
Traczewski MM, Carretto E, Canton R, Moore NM, Carba RST. Multicenter Evaluation of the Xpert Carba-R Assay for Detection of Carbapenemase Genes in Gram-Negative Isolates. J. Clin. Microbiol. 2018;56: e00272-18.
Moore NM, Canton R, Carretto E, et al. Rapid Identification of Five Classes of Carbapenem Resistance Genes Directly from Rectal Swabs by Use of the Xpert Carba-R Assay. J. Clin. Microbiol. 2017; 55:2268–2275.
Souverein D, Euser SM, van der Reijden WA, et al. Clinical sensitivity and specificity of the Check-Points Check-Direct ESBL Screen for BD MAX, a real-time AZMİT for direct ESBL detection from rectal swabs. J Antimicrob Chemother. 2017;72(9):2512-2518.
Huang TD, Bogaerts P, Ghilani E, et al. Multicentre evaluation of the Check-Direct CPE® assay for direct screening of carbapenemase-producing Enterobacteriaceae from rectal swabs. J Antimicrob Chemother. 2015;70(6):1669-73.
Lau AF, Fahle GA, Kemp MA, Jassem AN, Dekker JP, Frank KM. Clinical Performance of Check-Direct CPE, a Multiplex PCR for Direct Detection of bla(KPC), bla(NDM) and/or bla(VIM), and bla(OXA)-48 from Perirectal Swabs. J Clin Microbiol. 2015;53(12):3729-37.
Girlich D, Oueslati S, Bernabeu S, et al. Evaluation of the BD MAX Check-Points CPO Assay for the Detection of Carbapenemase Producers Directly from Rectal Swabs. J Mol Diagn. 2020;22(2):294-300.
Gonzalez C, Oueslati S, Biez L, Dortet L, Naas T. Evaluation of the EasyScreen™ ESBL/CPO Detection Kit for the Detection of ß-Lactam Resistance Genes. Diagnostics (Basel). 2022;12(9):2223.
Del Bianco F, Morotti M, Zannoli S et al. Comparison of Four Commercial Screening Assays for the Detection of blaKPC, blaNDM, blaIMP, blaVIM, and blaOXA48 in Rectal Secretion Collected by Swabs. Microorganisms. 2019;7(12):704.
Chen HY, Tseng HY, Chen CL, et al. The real-world impact of the BioFire FilmArray blood culture identification 2 panel on antimicrobial stewardship among patients with bloodstream infections in intensive care units with a high burden of drug-resistant pathogens. J Microbiol Immunol Infect. 2024;57(4):580-593.
Tojo M, Fujita T, Ainoda Y, et al. Evaluation of an automated rapid diagnostic assay for detection of Gram-negative bacteria and their drug-resistance genes in positive blood cultures. PLoS ONE. 2014;9:e94064.
Uno N, Suzuki H, Yamakawa H, et al. Multicenter evaluation of the Verigene Gram-negative blood culture nucleic acid test for rapid detection of bacteria and resistance determinants in positive blood cultures. Diagn Microbiol Infect Dis. 2015;83(4):344-8.
Kaprou GD, Bergšpica I, Alexa EA, Alvarez-Ordóñez A, Prieto M. Rapid Methods for Antimicrobial Resistance Diagnostics. Antibiotics (Basel). 2021;10(2):209.
Zou Y, Mason MG, Botella JR. Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics. PLoS ONE. 2020;15: e0235216.
García-Fernández S, Morosini MI, Marco F, et al. Evaluation of the eazyplex® SuperBug CRE system for rapid detection of carbapenemases and ESBLs in clinical Enterobacteriaceae isolates recovered at two Spanish hospitals. J Antimicrob Chemother. 2015;70(4):1047-50.
Srivastava P, Prasad D. Isothermal nucleic acid amplification and its uses in modern diagnostic technologies. 3 Biotech. 2023;13(6):200.
Sasano M, Seki M, Takano C, Komine-Aizawa S, Hayakawa S. An improved primer design for the loop-mediated isothermal amplification (LAMP) method to detect oxacillinase (OXA)-48 β-lactamase genes in Gram-negative bacteria for clinical applications. J Infect Chemother. 2021;27(7):1005-1012.
Seki M, Omagari D, Kilgore P, et al. Loop-Mediated Isothermal Amplification Assay for β-Lactamase Identification on Clinical Isolates of Pseudomonas aeruginosa. Open Forum Infect Dis. 2016;3(Suppl 1):181.
Carter B, Wu G, Woodward MJ, Anjum MF. A process for analysis of microarray comparative genomics hybridisation studies for bacterial genomes. BMC Genomics. 2008, 29;9:53.
Anjum MF, Zankari E, Hasman H. Molecular Methods for Detection of Antimicrobial Resistance. Microbiol Spectr. 2017;5(6): 10.1128/microbiolspec.arba-0011-2017.
Grimm V, Ezaki S, Susa M, Knabbe C, Schmid RD, Bachmann TT. Use of DNA microarrays for rapid genotyping of TEM ß-lactamases that confer resistance. J Clin Microbiol. 2004;42(8):3766-74.
Leinberger DM, Grimm V, Rubtsova M, et al. Integrated detection of extended-spectrum-beta-lactam resistance by DNA microarray-based genotyping of TEM, SHV, and CTX-M genes. J Clin Microbiol. 2010;48(2):460-71.
Bogaerts P, Hujer AM, Naas T, et al. Multicenter evaluation of a new DNA microarray for rapid detection of clinically relevant bla genes from beta-lactam-resistant gram-negative bacteria. Antimicrob Agents Chemother. 2011;55(9):4457-60.
Naas T, Cuzon G, Bogaerts P, Glupczynski Y, Nordmann P. Evaluation of a DNA microarray (Check-MDR CT102) for rapid detection of TEM, SHV, and CTX-M extended-spectrum ß-lactamases and of KPC, OXA-48, VIM, IMP, and NDM-1 carbapenemases. J Clin Microbiol. 2011;49:1608–1613.
Cuzon G, Naas T, Bogaerts P, Glupczynski Y, Nordmann P. Evaluation of a DNA microarray for the rapid detection of extended-spectrum ß-lactamases (TEM, SHV and CTX-M), plasmid-mediated cephalosporinases (CMY-2-like, DHA, FOX, ACC-1, ACT/MIR and CMY-1-like/MOX) and carbapenemases (KPC, OXA-48, VIM, IMP and NDM) J. Antimicrob. Chemother. 2012;67:1865–1869.
Braun SD, Jamil B, Syed MA, et al. Prevalence of carbapenemase-producing organisms at the Kidney Center of Rawalpindi (Pakistan) and evaluation of an advanced molecular microarray-based carbapenemase assay. Future Microbiol. 2018;13:1225-1246.
Rathmair F. (2023). Evaluation of the CarbDetect AS-2 microarray for the detection of resistance genes in Enterobacterales (Unpublished doctoral dissertation). Medical University of Vienna. https://repositorium.meduniwien.ac.at/obvumwhs/content/titleinfo/6208147.
Yamin D, Uskoković V, Wakil AM, et al. Current and Future Technologies for the Detection of Antibiotic-Resistant Bacteria. Diagnostics (Basel). 2023;13(20):3246.
Shin J, Kim SR, Xie Z, Jin YS, Wang YC. A CRISPR/Cas12a-Based System for Sensitive Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales. Biosensors (Basel). 2024;14(4):194.
Marcos DP, Fernández-Diego L, Rodríguez-Grande J, et al. An accurate amplification-free CRISPR/Cas12a-based assay for GES β-lactamase detection. Int J Antimicrob Agents. 2025;107506.
Yang JW, Kim H, Hyeon LS, Yoo JS, Kang S. Development of a Recombinase Polymerase Amplification-Coupled CRISPR/Cas12a Platform for Rapid Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales. Biosensors (Basel). 2024;14(11):536.
Florio W, Baldeschi L, Rizzato C, Tavanti A, Ghelardi E, Lupetti A. Detection of Antibiotic-Resistance by MALDI-TOF Mass Spectrometry: An Expanding Area. Front Cell Infect Microbiol. 2020;10:572909.
Burckhardt I, Zimmermann S. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours. J Clin Microbiol. 2011;49(9):3321-4.
Hrabák J, Chudácková E, Walková R. Matrix-assisted laser desorption ionization-time of flight (maldi-tof) mass spectrometry for detection of antibiotic resistance mechanisms: from research to routine diagnosis. Clin Microbiol Rev. 2013;26(1):103-14.
Oviano M, Sparbier K, Barba MJ, Kostrzewa M, Bou G. Universal protocol for the rapid automated detection of carbapenem-resistant gram-negative bacilli directly from blood cultures by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF/MS). Int. J. Antimicrob Agents, 2016;48: 655–660
Referanslar
Bush K, Jacoby GA. Updated functional classification of ß-lactamases. Antimicrob Agents Chemother. 2010;54(3):969-76.
Zhuang Q, Guo H, Peng T, et al. Advances in the detection of β-lactamase: A review. Int J Biol Macromol. 2023;251:126159.
Lawrence J, O'Hare D, van Batenburg-Sherwood J, Sutton M, Holmes A, Rawson TM. Innovative approaches in phenotypic ß-lactamase detection for personalised infection management. Nat Commun 2024; 15, 9070 https://doi.org/10.1038/s41467-024-53192-7
Elbehiry A, Marzouk E, Abalkhail A, et al. Detection of antimicrobial resistance via state-of-the-art technologies versus conventional methods. Front Microbiol. 2025;16:1549044.
Noster J, Thelen P, Hamprecht A. Detection of Multidrug-Resistant Enterobacterales-From ESBLs to Carbapenemases. Antibiotics (Basel). 2021 Sep 21;10(9):1140.
Priyanka Uprety, Thomas J Kirn. 2023. Molecular Detection of Antibacterial Drug Resistance In: Carroll KC, Pfaller MA Manual of Clinical Microbiology, 12th Edition. ASM Press, Washington, DC.(BU REF YAZIMI DOĞRU MU?)
Mullis K, Faloona F, Scharf S, Saiki R, Horn G, Erlich H. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb Symp Quant Biol. 1986;51(1):263-73.
Rahman M, Waseka AJ. Clinical Laboratory and Molecular Detection of Extended Spectrum ß lactamases: A Review Update. Bangladesh Journal of Infectious Diseases, 2015;1(1):12.
Sahoo R, Jadhav S, Nema V. Journey of technological advancements in the detection of antimicrobial resistance. J Formos Med Assoc. 2024 Apr;123(4):430-441.
Rood IGH, Li Q. Review: Molecular detection of extended spectrum-β-lactamase- and carbapenemase-producing Enterobacteriaceae in a clinical setting. Diagn Microbiol Infect Dis. 2017;89(3):245-250.
Zhuang Q, Guo H, Peng T, et al. Advances in the detection of β-lactamase: A review. Int J Biol Macromol. 2023;251:126159.
Traczewski MM, Carretto E, Canton R, Moore NM, Carba RST. Multicenter Evaluation of the Xpert Carba-R Assay for Detection of Carbapenemase Genes in Gram-Negative Isolates. J. Clin. Microbiol. 2018;56: e00272-18.
Moore NM, Canton R, Carretto E, et al. Rapid Identification of Five Classes of Carbapenem Resistance Genes Directly from Rectal Swabs by Use of the Xpert Carba-R Assay. J. Clin. Microbiol. 2017; 55:2268–2275.
Souverein D, Euser SM, van der Reijden WA, et al. Clinical sensitivity and specificity of the Check-Points Check-Direct ESBL Screen for BD MAX, a real-time AZMİT for direct ESBL detection from rectal swabs. J Antimicrob Chemother. 2017;72(9):2512-2518.
Huang TD, Bogaerts P, Ghilani E, et al. Multicentre evaluation of the Check-Direct CPE® assay for direct screening of carbapenemase-producing Enterobacteriaceae from rectal swabs. J Antimicrob Chemother. 2015;70(6):1669-73.
Lau AF, Fahle GA, Kemp MA, Jassem AN, Dekker JP, Frank KM. Clinical Performance of Check-Direct CPE, a Multiplex PCR for Direct Detection of bla(KPC), bla(NDM) and/or bla(VIM), and bla(OXA)-48 from Perirectal Swabs. J Clin Microbiol. 2015;53(12):3729-37.
Girlich D, Oueslati S, Bernabeu S, et al. Evaluation of the BD MAX Check-Points CPO Assay for the Detection of Carbapenemase Producers Directly from Rectal Swabs. J Mol Diagn. 2020;22(2):294-300.
Gonzalez C, Oueslati S, Biez L, Dortet L, Naas T. Evaluation of the EasyScreen™ ESBL/CPO Detection Kit for the Detection of ß-Lactam Resistance Genes. Diagnostics (Basel). 2022;12(9):2223.
Del Bianco F, Morotti M, Zannoli S et al. Comparison of Four Commercial Screening Assays for the Detection of blaKPC, blaNDM, blaIMP, blaVIM, and blaOXA48 in Rectal Secretion Collected by Swabs. Microorganisms. 2019;7(12):704.
Chen HY, Tseng HY, Chen CL, et al. The real-world impact of the BioFire FilmArray blood culture identification 2 panel on antimicrobial stewardship among patients with bloodstream infections in intensive care units with a high burden of drug-resistant pathogens. J Microbiol Immunol Infect. 2024;57(4):580-593.
Tojo M, Fujita T, Ainoda Y, et al. Evaluation of an automated rapid diagnostic assay for detection of Gram-negative bacteria and their drug-resistance genes in positive blood cultures. PLoS ONE. 2014;9:e94064.
Uno N, Suzuki H, Yamakawa H, et al. Multicenter evaluation of the Verigene Gram-negative blood culture nucleic acid test for rapid detection of bacteria and resistance determinants in positive blood cultures. Diagn Microbiol Infect Dis. 2015;83(4):344-8.
Kaprou GD, Bergšpica I, Alexa EA, Alvarez-Ordóñez A, Prieto M. Rapid Methods for Antimicrobial Resistance Diagnostics. Antibiotics (Basel). 2021;10(2):209.
Zou Y, Mason MG, Botella JR. Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics. PLoS ONE. 2020;15: e0235216.
García-Fernández S, Morosini MI, Marco F, et al. Evaluation of the eazyplex® SuperBug CRE system for rapid detection of carbapenemases and ESBLs in clinical Enterobacteriaceae isolates recovered at two Spanish hospitals. J Antimicrob Chemother. 2015;70(4):1047-50.
Srivastava P, Prasad D. Isothermal nucleic acid amplification and its uses in modern diagnostic technologies. 3 Biotech. 2023;13(6):200.
Sasano M, Seki M, Takano C, Komine-Aizawa S, Hayakawa S. An improved primer design for the loop-mediated isothermal amplification (LAMP) method to detect oxacillinase (OXA)-48 β-lactamase genes in Gram-negative bacteria for clinical applications. J Infect Chemother. 2021;27(7):1005-1012.
Seki M, Omagari D, Kilgore P, et al. Loop-Mediated Isothermal Amplification Assay for β-Lactamase Identification on Clinical Isolates of Pseudomonas aeruginosa. Open Forum Infect Dis. 2016;3(Suppl 1):181.
Carter B, Wu G, Woodward MJ, Anjum MF. A process for analysis of microarray comparative genomics hybridisation studies for bacterial genomes. BMC Genomics. 2008, 29;9:53.
Anjum MF, Zankari E, Hasman H. Molecular Methods for Detection of Antimicrobial Resistance. Microbiol Spectr. 2017;5(6): 10.1128/microbiolspec.arba-0011-2017.
Grimm V, Ezaki S, Susa M, Knabbe C, Schmid RD, Bachmann TT. Use of DNA microarrays for rapid genotyping of TEM ß-lactamases that confer resistance. J Clin Microbiol. 2004;42(8):3766-74.
Leinberger DM, Grimm V, Rubtsova M, et al. Integrated detection of extended-spectrum-beta-lactam resistance by DNA microarray-based genotyping of TEM, SHV, and CTX-M genes. J Clin Microbiol. 2010;48(2):460-71.
Bogaerts P, Hujer AM, Naas T, et al. Multicenter evaluation of a new DNA microarray for rapid detection of clinically relevant bla genes from beta-lactam-resistant gram-negative bacteria. Antimicrob Agents Chemother. 2011;55(9):4457-60.
Naas T, Cuzon G, Bogaerts P, Glupczynski Y, Nordmann P. Evaluation of a DNA microarray (Check-MDR CT102) for rapid detection of TEM, SHV, and CTX-M extended-spectrum ß-lactamases and of KPC, OXA-48, VIM, IMP, and NDM-1 carbapenemases. J Clin Microbiol. 2011;49:1608–1613.
Cuzon G, Naas T, Bogaerts P, Glupczynski Y, Nordmann P. Evaluation of a DNA microarray for the rapid detection of extended-spectrum ß-lactamases (TEM, SHV and CTX-M), plasmid-mediated cephalosporinases (CMY-2-like, DHA, FOX, ACC-1, ACT/MIR and CMY-1-like/MOX) and carbapenemases (KPC, OXA-48, VIM, IMP and NDM) J. Antimicrob. Chemother. 2012;67:1865–1869.
Braun SD, Jamil B, Syed MA, et al. Prevalence of carbapenemase-producing organisms at the Kidney Center of Rawalpindi (Pakistan) and evaluation of an advanced molecular microarray-based carbapenemase assay. Future Microbiol. 2018;13:1225-1246.
Rathmair F. (2023). Evaluation of the CarbDetect AS-2 microarray for the detection of resistance genes in Enterobacterales (Unpublished doctoral dissertation). Medical University of Vienna. https://repositorium.meduniwien.ac.at/obvumwhs/content/titleinfo/6208147.
Yamin D, Uskoković V, Wakil AM, et al. Current and Future Technologies for the Detection of Antibiotic-Resistant Bacteria. Diagnostics (Basel). 2023;13(20):3246.
Shin J, Kim SR, Xie Z, Jin YS, Wang YC. A CRISPR/Cas12a-Based System for Sensitive Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales. Biosensors (Basel). 2024;14(4):194.
Marcos DP, Fernández-Diego L, Rodríguez-Grande J, et al. An accurate amplification-free CRISPR/Cas12a-based assay for GES β-lactamase detection. Int J Antimicrob Agents. 2025;107506.
Yang JW, Kim H, Hyeon LS, Yoo JS, Kang S. Development of a Recombinase Polymerase Amplification-Coupled CRISPR/Cas12a Platform for Rapid Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales. Biosensors (Basel). 2024;14(11):536.
Florio W, Baldeschi L, Rizzato C, Tavanti A, Ghelardi E, Lupetti A. Detection of Antibiotic-Resistance by MALDI-TOF Mass Spectrometry: An Expanding Area. Front Cell Infect Microbiol. 2020;10:572909.
Burckhardt I, Zimmermann S. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours. J Clin Microbiol. 2011;49(9):3321-4.
Hrabák J, Chudácková E, Walková R. Matrix-assisted laser desorption ionization-time of flight (maldi-tof) mass spectrometry for detection of antibiotic resistance mechanisms: from research to routine diagnosis. Clin Microbiol Rev. 2013;26(1):103-14.
Oviano M, Sparbier K, Barba MJ, Kostrzewa M, Bou G. Universal protocol for the rapid automated detection of carbapenem-resistant gram-negative bacilli directly from blood cultures by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF/MS). Int. J. Antimicrob Agents, 2016;48: 655–660
