TY - JOUR T1 - Comparison of the Modified Hodge Test and Inhibitor-Based Tests for Detection of Carbapenemase-Producing Enterobacteriaceae Isolates TT - Karbapenemaz Üreticisi Enterobacteriaceae İzolatlarının Saptanmasında Modifiye Hodge Testi ile İnhibitör Tabanlı Testlerin Karşılaştırılması AU - Us, Ebru PY - 2016 DA - December JF - Ankara Üniversitesi Tıp Fakültesi Mecmuası PB - Ankara University WT - DergiPark SN - 0365-8104 SP - 152 EP - 157 VL - 69 IS - 3 LA - en AB - Aim: Rapid and accurate detection of carbapenemase producing Enterobacteriaceae isolates plays an important role in preventing the spread and defining treatment strategies.The modified Hodge test (MHT) is an easy and inexpensive tool that can be used to detect carbapenemase activity. Another method used for the same purpose are inhibitor-based tests (IBTs). The aim of this study is to compare the performances of these two phenotypic methods in the detection of carbapenemases in Enterobacteriaceae isolates.Materials and Methods: A total of 112 non-repeat Enterobacteriaceae strains isolated from various clinical samples at Ibni Sina Hospital Central Microbiology Laboratory between 2010-2014 that showed decreased sensitivity and resistance to ertapenem by Kirby-Bauer disk diffusion method were included in the study.Results: When 1:10 dilution [0,05 McFarland (McF)] was performed, out of 94 isolates positive for carbapenemase-encoding genes (CEGs), false negativity was detected in seven isolates, whereas out of 18 isolates negative for CEGs, false positivity was detected in 16 isolates; so the sensitivity of MHT for carbapenemase detection was found to be 92,6%. Without dilution (0.5 McF) the number of false negative isolates decreased to three and the sensitivity of MHT increased to 96,8%, (p KW - Enterobacteriaceae KW - Carbapenemase KW - Modified Hodge Test KW - İnhibitor-Based Tests N2 - Amaç: Karbapenemaz üreten Enterobacteriaceae izolatlarının hızlı ve doğru tanısı, yayılımı önleme ve tedavi stratejilerini belirlemede önemlidir. Modifiye Hodge testi (MHT) karbapenemaz aktivitesini fenotipik olarak saptamak için kullanılan kolay ve ucuz bir yöntemdir. Aynı amaçla kullanılan diğer yöntem inhibitör tabanlı testlerdir (İTT). Bu çalışmanın amacı bu iki fenotipik yöntemin Enterobacteriaceae izolatlarında karbapenemazları saptama performanslarının karşılaştırılmasıdır.Gereç ve Yöntem: 2010-2014 yılları arasında Ankara Üniversitesi Tıp Fakültesi İbni Sina Hastanesi Merkez Mikrobiyoloji Laboratuvarı’nda farklı hastalara ait çeşitli klinik örneklerden izole edilen, Kirby-Bauer disk difüzyon yöntemiyle ertapeneme orta duyarlılık ya da direnç gösteren Enterobacteriaceae ailesindeki 112 adet tekrar etmeyen klinik izolat çalışmaya dahil edilmiştir.Bulgular: MHT, 1:10 sulandırım [0,05 McFarland (McF)] kullanılarak yapıldığında karbapenemaz kodlayan gen (KKG)’leri pozitif 94 izolatın 7’sinde yalancı negatif; KKG negatif 18 izolatın 16’sında yalancı pozitif sonuç alınmış ve MHT’nin karbapenemazları saptamadaki duyarlılığı % 92,6 bulunmuştur. Sulandırım adımı uygulanmadığında (0,5 McF) yalancı negatif izolat sayısı 3’e düşmüş ve testin duyarlılığı % 96,8’e yükselmiştir (p CR - 1. Nordmann P, Cornaglia G. Carbapenemaseproducing Enterobacteriaceae: a call for action! Clin Microbiol Infect 2012; 18:411-412. CR - 2. Budak S, Aktaş Z, Erdem H. Enterik gramnegatif bakterilerde laboratuvardan kliniğe karbapenemazlar. Mediterr J Infect Microbes Antimicrob 2012; 1:1-11. CR - 3. Walsh TR. Emerging carbapenemases: a global perspective. Int J of Antimicrob Agents 2010; 36:S8-14. CR - 4. Nordmann P, Poirel L. Strategies for identification of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother 2013; 68:487-489. CR - 5. Hammoudi D, Moubareck CA, Sarkis DK. How to detect carbapenemase producers? A literature review of phenotypic and molecular methods. J Microbiol Methods 2014; 107:106-118. CR - 6. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. Twenty-fifth Informational Supplement. CLSI Document M100- S25, 2015. CLSI, Wayne, PA. CR - 7. Girlich D, Poirel L, Nordmann P. Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol 2012; 50:477-479. CR - 8. European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 5.0. Available at: http://www.eucast.org/clinical_breakpoints/ CR - 9. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. Twenty-fourth Informational Supplement. CLSI Document M100-S24, 2014. CLSI, Wayne, PA. CR - 10. Poirel L, Walsh TR, Cuvillier V, et al. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011; 70:119-123. CR - 11. Kutlu HH. Çeşitli Klinik Örneklerden İzole Edilen Gram Negatif Enterik Bakterilerde Karbapenemaz Varlığının ve Tiplerinin Araştırılması. Ankara Üniversitesi Tıp Fakültesi Tıbbi Mikrobiyoloji Anabilim Dalı Tıpta Uzmanlık Tezi. Ankara. 2015 CR - 12. Lee K, Kim CK, Yong D, et al. Improved performance of the modified Hodge test with MacConkey agar for screening carbapenemase-producing Gram-negative bacilli. J Microbiol Methods 2010; 83:149-152. CR - 13. Liofilchem® KPC&MBL&OXA-48 disc kit (acc. to EUCAST). Available at: http://www.liofilchem.net/login/pd/pi/99007_PI.pdf CR - 14. Coudron PE. Inhibitor-Based Methods for Detection of Plasmid-Mediated AmpC β-Lactamases in Klebsiella spp., Escherichia coli, and Proteus mirabilis. J Clin Microbiol 2005; 43:4163-4167. CR - 15. Kim H, Park JS, Sung H, et al. Further Modification of the Modified Hodge Test for Detecting Metallo-β-Lactamase-Producing Carbapenem-Resistant Enterobacteriaceae. Ann Lab Med 2015; 35:298-305. CR - 16. Miriagou V, Cornaglia G, Edelstein M, et al. Acquired carbapenemases in Gram-negative bacterial pathogens: detection and surveillance issues. Clin Microbiol Infect 2010; 16:112-122. CR - 17. Cohen Stuart J, Leverstein-Van Hall MA. Dutch Working Party on the Detection of Highly Resistant Microorganisms. Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrob Agents 2010; 36:205-210. CR - 18. Eser OK, Altun Uludağ H, Ergin A, ve ark. İnvazif enfeksiyonlara neden olan GSBL pozitif Enterobacteriaceae izolatlarında karbapenem direnci. Mikrobiyol Bul 2014; 48:59-69. CR - 19. Van Dijk K, Voets GM, Scharringa J, et al. A disc diffusion assay for detection of class A, B and OXA-48 carbapenemases in Enterobacteriaceae using phenyl boronic acid, dipicolinic acid and temocillin. Clin Microbiol Infect 2014; 20:345-349. CR - 20. Saito R, Koyano S, Dorin M, et al. Evaluation of a simple phenotypic method for the detection of carbapenemase-producing Enterobacteriaceae. J Microbiol Methods 2015; 108:45-48. CR - 21. Bayramoğlu G, Uluçam G, Gençoğlu Özgür Ç, ve ark. Enterobacteriaceae izolatlarında karbapenemazların saptanmasında modifiye Hodge testi ve Carba NP testlerinin karşılaştırılması. Mikrobiyol Bul 2016; 50:1-10. CR - 22. Lutring JD, Limbago BM. The problem of carbapenemase producing carbapenem-resistantEnterobacteriaceae detection.J Clin Microbiol 2016; doi:10.1128/JCM.02771-15. CR - 23. Tsakris A, Poulou A, Pournaras S, et al. A simple phenotypic method for the differentiation of metallo-beta-lactamases and class A KPC carbapenemases in Enterobacteriaceae clinical isolates. J Antimicrob Chemother 2010; 65:1664-1671. CR - 24. Seah C, Low DE, Patel SN, et al. Comparative evaluation of a chromogenic agar edium, the modified Hodge test, and a battery of meropenem-inhibitor discs for detection of carbapenemase activity in Enterobacteriaceae. J Clin Microbiol 2011; 49:1965-1969. CR - 25. Huang TD, Poirel L, Bogaerts P, et al. Temocillin and piperacillin/tazobactam resistance by disc diffusion as antimicrobial surrogate markers for the detection of carbapenemase-producing Enterobacteriaceae in geographical areas with a high prevalence of OXA-48 producers. J Antimicrob Chemother 2014; 69: 445-450. CR - 26. Hansen F, Hammerum AM, Skov RL, et al. Evaluation of Rosco Neo-Sensitabs for phenotypic detection and subgrouping of ESBL-, AmpC- and carbapenemase-producing Enterobacteriaceae. APMIS 2012; 120:724-732. CR - 27. Giske CG, Gezelius L, Samuelsen Ø, et al. A sensitive and specific phenotypic assay for detection of metallo-β-lactamases and KPC in Klebsiella pneumoniae with the use of meropenem disks supplemented with aminophenylboronic acid, dipicolinic acid and cloxacillin. Clin Microbiol Infect 2011; 17:552-556. CR - 28. Labarca J, Poirel L, Ozdamar M, et al. KPC-producing Klebsiella pneumoniae, finally targeting Turkey. New Microbes New Infect 2014; 2:50-51. CR - 29. Demir Y, Zer Y, Karaoglan I. Investigation of VIM, IMP, NDM-1, KPC AND OXA48 enzymes in Enterobacteriaceae strains. Pak J Pharm Sci 2015; 28(3 Suppl):1127-1133 CR - 30. Sahin K, Tekin A, Ozdas S, et al. Evaluation of carbapenem resistance using phenotypic and genotypic techniques in Enterobacteriaceae isolates. Ann Clin Microbiol Antimicrob 2015; 14:44. CR - 31. Iraz M, Özad Düzgün A, Sandallı C, et al. Distribution of β-lactamase genes among carbapenem-resistant Klebsiella pneumoniae strains isolated from patients in Turkey. Ann Lab Med 2015; 35: 595-601. UR - https://dergipark.org.tr/en/pub/autfm/issue//1717201 L1 - https://dergipark.org.tr/en/download/article-file/4948553 ER -