Genişlemiş Spektrumlu β-laktamazların Saptanmasında Kullanılan Fenotipik Testler
Özet
Genişlemiş spektrumlu β-laktamaz (GSBL) üreten Enterobacterales izolatlarının saptanması, antibiyotik tedavi yönetimi ve enfeksiyon kontrolü açısından, klinik mikrobiyoloji laboratuvarının önemli bir rolüdür. GSBL saptanmasında, çift disk sinerji testi (ÇDST), GSBL gradiyent şerit testi, mikrodilüsyon ve kombinasyon disk yöntemi gibi, üçüncü kuşak bir sefalosporin ve klavulanat arasındaki sinerjiye dayanan çeşitli fenotipik testler tasarlanmıştır. Bu derlemede, ‘’The European Committee on Antimicrobial Susceptibility Testing’’ (EUCAST) önerilerine göre GSBL saptanması için kullanılan fenotipik yöntemler açıklanmaktadır.
Detection of an extended spectrum β-lactamase producing Enterobacterales, is an important role of the clinical microbiology laboratory regarding both antimicrobial treatment and infection control. Several phenotypic ESBL detection tests, based on the synergy between a third-generation cephalosporin and clavulanate, have been designed: the double-disk synergy test (DDST), ESBL gradient test, microdilution test, and the combination disk method. This review describes the phenotypic methods for ESBL detection according to The European Committee on Antimicrobial Susceptibility Testing (EUCAST) suggestions.
Referanslar
Rahman SU, Ali T, Ali I, et al. The Growing Genetic and Functional Diversity of Extended Spectrum β-Lactamases. BioMed Res Int. 2018;2018(26):9519718. doi: 10.1155/2018/9519718.
Pana ZD, Zaoutis T. Treatment of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBLs) infections: What have we learned until now? Faculty Rev. 2018;29(7):F1000. doi: 10.12688/f1000research.14822.1.
Mulani MS, Kamble EE, Kumkar SN, et al. Emerging Strategies to Combat ESKAPE Pathogens in the Era of Antimicrobial Resistance: A Review. Front Microbiol. 2019;4(1):539. doi: 10.3389/fmicb.2019.00539.
Castanheira M, Simner PJ, Bradford PA. Extended-spectrum-lactamases: An update on their characteristics, epidemiology and detection. JAC Antimicrob Resist. 2021;16(3):dlab092. doi: 10.1093/jacamr/dlab092.
Bush K, Jacoby GA. Medeiros, A.A. A functional classification scheme for β-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother. 1995;39(6):1211-33. doi: 10.1128/AAC.39.6.1211.
Bush K, Jacoby GA. Updated functional classification of β-lactamases. Antimicrob Agents Chemother. 2010;54(3):969-76. doi:10.1128/AAC.01009-09.
http://bldb.eu/Enzymes.php. Last updated: May 19, 2025.
Husna A, Rahman MM, Badruzzaman ATM, et al. Extended-Spectrum-Lactamases (ESBL): challenges and opportunities. Biomedicines 2023;11(11);2937. doi:10.3390/biomedicines11112937.
Guidelines for Detection of Resistance Mechanisms and Specific Resistance of Clinical and/or Epidemiological Importance, Version 2.0, 2017. https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Resistance_mechanisms/EUCAST_detection_of_resistance_mechanisms_170711.pdf.
Bush K, Bradford PA. Epidemiology of b-lactamase-producing pathogens. Clin Microbiol Rev. 2020; 33(2):e00047-19. doi:10.1128/CMR.00047-19.
El-JadeMR, ParcinaM, Schmithausen RMet al. ESBL detection: comparisonof a commercially available chromogenic test for third generationcephalosporine resistance and automated susceptibility testing in Enterobactericeae. PLoS One 2016 ;11(8):e0160203. doi:10.1371/journal.pone.0160203.
Thomson KS, Cornish NE, Hong SG et al. Comparison of phoenix and VITEK 2 Extended-Spectrum-b-Lactamase Detection Tests for Analysis of Escherichia coli and Klebsiella isolates with well-characterized β-lactamases. J Clin Microbiol. 2007;45(8):2380-4. doi:10.1128/JCM.00776-07.
Drieux L, Brossier F, Sougakoff W, et al. Phenotypic detection of extended-spectrum β-lactamase production in Enterobacteriaceae: review and bench guide. Clin Microbiol Infect. 2008;14(1):90-103. doi:10.1111/j.1469-0691.2007.01846.x.
Evulation of a new cefepime-clavulanate ESBL Etest to detect extended-spectrum β-lactamases in a Enterobacteriaceae starin cellection. J Antimicrob Chemother. 2004;54(1):134-8. doi:10.1093/jac/dkh274.
Wiegand I, Geiss HK, Mack D, et al. Detection of extended-spectrum β-lactamases among Enterobacteriaceae by use of semiautomated microbiology systems and manual detection procedures. J Clin Microbiol. 2007;45(4):1167-74. doi:10.1128/JCM.01988-06.
Sanguinetti M, Posteraro B, SpanuT, et al. Characterization of clinical isolates of Enterobacteriaceae from Italy by the BD Phoenix extended-spectrum β-lactamase detection method. J Clin Microbiol. 2003;41(4):1463-8. doi:10.1128/JCM.41.4.1463-1468.2003.
Champs C, Monne C, Bonnet R, et al. New TEM variant (TEM92) produced by Proteus mirabilis and Providencia stuartii isolates. Antimicrob Agents Chemother. 2001;45(4):1278-80. doi:10.1128/AAC.45.4.1278-1280.2001.
Balko T, Karlowsky JA, Palatnick LP, et al. Cracterization of the inoculum effect with Haemophilus influenzae and b-lactams. Diagn Microbiol Infect Dis. 1999;33(1):47-58. doi:10.1016/s0732-8893(98)00117-5.
Sykes RB, Matthew M. The β-lactamases of gram-negative bacteria and their role in resistance to b-lactam antibiotics. J Antimicrob Chemother. 1976;2(2):115-57. doi:10.1093/jac/2.2.115.
Soriano F, Santamaria M, Ponte C, et al. In vivo significance of the inoculum effect of antibiotics on Escherichia coli. Eur J Clin Microbiol Infect Dis. 1988;7(3):410-2. Doi:10.1007/BF01962350.
Queenan AM, Foleno B, Gownley C, et al. Effects of inoculum and b-lactamase activity in AmpC- and extended-spectrum blactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniaeclinical isolates tested by using NCCLS ESBL methodology. J Clin Microbiol. 2004;42(1):269-75. doi:10.1128/JCM.42.1.269-275.2004.
Burgess DS, Hall RG. In vitro killing of parenteral b-lactams againststandard and high inocula of extended-spectrum b-lactamase and nonESBL producing Klebsiella pneumoniae. Diagn Microbiol Infect Dis. 2004;49(1):41-6. doi:10.1016/j.diagmicrobio.2003.11.007.
Craig WA, Bhavnani SM, Ambrose PG. The inoculum effect: factor or artifact? Diagn Microbiol Infect Dis. 2004;50(4):229-30. doi:10.1016/j.diagmicrobio.2004.07.006.
Ramphal R, Ambrose PG. Extended-spectrum b-lactamases and clinical outcomes: current data. Clin Infect Dis 2006;42(Suppl 4):S164–72. doi:10.1086/500663.
Komatsu M, Aihara M, Shimakawa K, et al. Evaluation of MicroScanESBL confirmation panel for Enterobacteriaceae-producing, extended spectrum β-lactamases isolated in Japan. Diagn Microbiol Infect Dis. 2003;46(2):125-30. doi:10.1016/s0732-8893(03)00041-5.
Referanslar
Rahman SU, Ali T, Ali I, et al. The Growing Genetic and Functional Diversity of Extended Spectrum β-Lactamases. BioMed Res Int. 2018;2018(26):9519718. doi: 10.1155/2018/9519718.
Pana ZD, Zaoutis T. Treatment of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBLs) infections: What have we learned until now? Faculty Rev. 2018;29(7):F1000. doi: 10.12688/f1000research.14822.1.
Mulani MS, Kamble EE, Kumkar SN, et al. Emerging Strategies to Combat ESKAPE Pathogens in the Era of Antimicrobial Resistance: A Review. Front Microbiol. 2019;4(1):539. doi: 10.3389/fmicb.2019.00539.
Castanheira M, Simner PJ, Bradford PA. Extended-spectrum-lactamases: An update on their characteristics, epidemiology and detection. JAC Antimicrob Resist. 2021;16(3):dlab092. doi: 10.1093/jacamr/dlab092.
Bush K, Jacoby GA. Medeiros, A.A. A functional classification scheme for β-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother. 1995;39(6):1211-33. doi: 10.1128/AAC.39.6.1211.
Bush K, Jacoby GA. Updated functional classification of β-lactamases. Antimicrob Agents Chemother. 2010;54(3):969-76. doi:10.1128/AAC.01009-09.
http://bldb.eu/Enzymes.php. Last updated: May 19, 2025.
Husna A, Rahman MM, Badruzzaman ATM, et al. Extended-Spectrum-Lactamases (ESBL): challenges and opportunities. Biomedicines 2023;11(11);2937. doi:10.3390/biomedicines11112937.
Guidelines for Detection of Resistance Mechanisms and Specific Resistance of Clinical and/or Epidemiological Importance, Version 2.0, 2017. https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Resistance_mechanisms/EUCAST_detection_of_resistance_mechanisms_170711.pdf.
Bush K, Bradford PA. Epidemiology of b-lactamase-producing pathogens. Clin Microbiol Rev. 2020; 33(2):e00047-19. doi:10.1128/CMR.00047-19.
El-JadeMR, ParcinaM, Schmithausen RMet al. ESBL detection: comparisonof a commercially available chromogenic test for third generationcephalosporine resistance and automated susceptibility testing in Enterobactericeae. PLoS One 2016 ;11(8):e0160203. doi:10.1371/journal.pone.0160203.
Thomson KS, Cornish NE, Hong SG et al. Comparison of phoenix and VITEK 2 Extended-Spectrum-b-Lactamase Detection Tests for Analysis of Escherichia coli and Klebsiella isolates with well-characterized β-lactamases. J Clin Microbiol. 2007;45(8):2380-4. doi:10.1128/JCM.00776-07.
Drieux L, Brossier F, Sougakoff W, et al. Phenotypic detection of extended-spectrum β-lactamase production in Enterobacteriaceae: review and bench guide. Clin Microbiol Infect. 2008;14(1):90-103. doi:10.1111/j.1469-0691.2007.01846.x.
Evulation of a new cefepime-clavulanate ESBL Etest to detect extended-spectrum β-lactamases in a Enterobacteriaceae starin cellection. J Antimicrob Chemother. 2004;54(1):134-8. doi:10.1093/jac/dkh274.
Wiegand I, Geiss HK, Mack D, et al. Detection of extended-spectrum β-lactamases among Enterobacteriaceae by use of semiautomated microbiology systems and manual detection procedures. J Clin Microbiol. 2007;45(4):1167-74. doi:10.1128/JCM.01988-06.
Sanguinetti M, Posteraro B, SpanuT, et al. Characterization of clinical isolates of Enterobacteriaceae from Italy by the BD Phoenix extended-spectrum β-lactamase detection method. J Clin Microbiol. 2003;41(4):1463-8. doi:10.1128/JCM.41.4.1463-1468.2003.
Champs C, Monne C, Bonnet R, et al. New TEM variant (TEM92) produced by Proteus mirabilis and Providencia stuartii isolates. Antimicrob Agents Chemother. 2001;45(4):1278-80. doi:10.1128/AAC.45.4.1278-1280.2001.
Balko T, Karlowsky JA, Palatnick LP, et al. Cracterization of the inoculum effect with Haemophilus influenzae and b-lactams. Diagn Microbiol Infect Dis. 1999;33(1):47-58. doi:10.1016/s0732-8893(98)00117-5.
Sykes RB, Matthew M. The β-lactamases of gram-negative bacteria and their role in resistance to b-lactam antibiotics. J Antimicrob Chemother. 1976;2(2):115-57. doi:10.1093/jac/2.2.115.
Soriano F, Santamaria M, Ponte C, et al. In vivo significance of the inoculum effect of antibiotics on Escherichia coli. Eur J Clin Microbiol Infect Dis. 1988;7(3):410-2. Doi:10.1007/BF01962350.
Queenan AM, Foleno B, Gownley C, et al. Effects of inoculum and b-lactamase activity in AmpC- and extended-spectrum blactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniaeclinical isolates tested by using NCCLS ESBL methodology. J Clin Microbiol. 2004;42(1):269-75. doi:10.1128/JCM.42.1.269-275.2004.
Burgess DS, Hall RG. In vitro killing of parenteral b-lactams againststandard and high inocula of extended-spectrum b-lactamase and nonESBL producing Klebsiella pneumoniae. Diagn Microbiol Infect Dis. 2004;49(1):41-6. doi:10.1016/j.diagmicrobio.2003.11.007.
Craig WA, Bhavnani SM, Ambrose PG. The inoculum effect: factor or artifact? Diagn Microbiol Infect Dis. 2004;50(4):229-30. doi:10.1016/j.diagmicrobio.2004.07.006.
Ramphal R, Ambrose PG. Extended-spectrum b-lactamases and clinical outcomes: current data. Clin Infect Dis 2006;42(Suppl 4):S164–72. doi:10.1086/500663.
Komatsu M, Aihara M, Shimakawa K, et al. Evaluation of MicroScanESBL confirmation panel for Enterobacteriaceae-producing, extended spectrum β-lactamases isolated in Japan. Diagn Microbiol Infect Dis. 2003;46(2):125-30. doi:10.1016/s0732-8893(03)00041-5.
