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Evaluation of the commercial combined disk test and minimum inhibitory concentration (MIC) determination for detection of carbapenemase producers among Gram-negative bacilli isolated in a region with high prevalence of blaOXA-48 and blaNDM

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Abstract

Carbapenem-resistant Gram-negative bacilli (GNB) are a concern in the Middle East and worldwide. Simple screening methods have been sought to detect carbapenemase producers to determine appropriate therapeutic measures and implement infection control interventions. In this study, we evaluated the efficiency of agar disc diffusion, commercial combined disc test (Rosco), and carbapenem MIC determination in comparison to molecular detection of carbapenemase genes among 82 carbapenem non-susceptible Enterobacteriaceae (CNSE) and 37 Acinetobacter/Pseudomonas isolates. The blaOXA-48, blaNDM, blaNDM/OXA-48, and blaIMP were detected in 68 out of 82 CNSE isolates. All of the Acinetobacter baumannii isolates were positive for the blaOXA-51 (n = 23), of those some were positive for blaOXA-48 (n = 13) and blaNDM (n = 3). Sensitivities and specificities of combined disc test for detection of blaNDM and blaOXA-48 carrying Enterobacteriaceae isolates were 92.5% and 100%, and 58.5% and 100%, respectively, while those for Acinetobacter/Pseudomonas isolates were 100%, 81.8% and 96.2%, 89%, respectively. While carbapenem MIC values had excellent concordance with phenotypic combined disc test for detection of blaOXA-48 producers (area under curve > 90%), only ertapenem MIC’s could precisely detect blaOXA-48 PCR-positive Enterobacteriaceae isolates (AUC 70%, sensitivity 70%, specificity 50%). The phenotypic commercial test showed excellent sensitivity for detection of blaNDM producers, but had poor sensitivity for blaOXA-48-producing Enterobacteriaceae. Ertapenem MIC values had low sensitivity and specificity for detection of the blaOXA-48-carrying Enterobacteriaceae. This is the first report of A. baumannii isolates co-harbored the blaOXA-48/blaNDM carbapenemases from Iran.

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References

  • Bartolini A, Frasson I, Cavallaro A, Richter SN, Palù G (2014) Comparison of phenotypic methods for the detection of carbapenem non susceptible Enterobacteriaceae. Gut Pathog 6:13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bharadwaj R, Joshi S, Dohe V, Gaikwad V, Kulkarni G, Shouche Y (2012) Prevalence of New Delhi metallo-β-lactamase (NDM-1)-positive bacteria in a tertiary care centre in Pune, India. Int J Antimicrob Agents 39:265–266

    Article  CAS  PubMed  Google Scholar 

  • Biglari S, Hanafiah A, Mohd Puzi S, Ramli R, Rahman M, Lopes BS (2016) Antimicrobial resistance mechanisms and genetic diversity of multidrug–resistant Acinetobacter baumannii isolated from a teaching hospital in Malaysia. Microb Drug Resist 23:545–555

    Article  CAS  PubMed  Google Scholar 

  • Bogaerts P, Rezende de Castro R, de Mendonça R, Huang TD, Denis O, Glupczynski Y (2013) Validation of carbapenemase and extended-spectrum b-lactamase multiplex endpoint PCR assays according to ISO 15189. J Antimicrob Chemother 68:1576–1582

    Article  CAS  PubMed  Google Scholar 

  • Brink AJ, Coetzee J, Corcoran C, Clay CG, Hari-Makkan D, Jacobson RK (2013) Emergence of OXA-48 and OXA-181 carbapenemases among Enterobacteriaceae in South Africa and evidence of in vivo selection of colistin resistance as a consequence of selective decontamination of the gastrointestinal tract. J Clin Microbiol 51:369–372

    Article  PubMed  PubMed Central  Google Scholar 

  • Clinical and Laboratory Standards Institute (CLSI) (2016) Performance standards for antimicrobial susceptibility testing; twenty-second informational supplements. CLSI document M100S-S26

  • Dortet L, Poirel L, Nordmann P (2014) Worldwide dissemination of the NDM-type carbapenemases in gram-negative bacteria. Biomed Res Int 249856. https://doi.org/10.1155/2014/249856

  • Dortet L, Bernabeu S, Gonzalez C, Naas T (2018) Evaluation of the carbapenem detection set™ for the detection and characterization of carbapenemase-producing Enterobacteriaceae. Diagn Microbiol Infect Dis 91:220–225

    Article  CAS  PubMed  Google Scholar 

  • Elsherif R, Ismail D, Elawady S, Jastaniah S, Al-Masaudi S, Harakeh S (2016) Boronic acid disk diffusion for the phenotypic detection of polymerase chain reaction-confirmed, carbapenem-resistant, gram-negative bacilli isolates. BMC Microbiol 16:135–140

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Endimiani A, Paterson DL (2007) Optimizing therapy for infections caused by Enterobacteriaceae producing extended-spectrum beta-lactamases. Semin Respir Crit Care Med 28:646–655

    Article  PubMed  Google Scholar 

  • Girlich D, Poirel L, Nordmann p (2012) Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol 50:477–479

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hall GS (1995) Enterobacteriaceae. In: Mahon CR, LDMG (eds) Textbook of diagnostic microbiology, 3rd edn. Saunders Company, New York, pp 464–514

    Google Scholar 

  • Hojabri Z, Ahangarzadeh Rezaee M, Nahaei MR, Soroush MH, Ghojazadeh M (2013) Comparison of in vitro activity of doripenem versus old carbapenems against Pseudomonas aeruginosa clinical isolates from both CF and burn patients. Adv Pharm Bull 3:121–125

    PubMed  PubMed Central  Google Scholar 

  • Hojabri Z, Pajand O, Bonura C, Aleo A, Giammanco A, Mammina C (2014) Molecular epidemiology of Acinetobacter baumannii in Iran: endemic and epidemic spread of multiresistant isolates. J Antimicrob Chemother 69:2383–2387

    Article  CAS  PubMed  Google Scholar 

  • Hrabák J, Chudáčková E, Papagiannitsis CC (2014) Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories. Clin Microbiol Infect 20:839–853

    Article  PubMed  Google Scholar 

  • Huang TD, Poirel L, Bogaerts P, Berhin C, Nordmann P, Glupczynski Y (2014) 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 69:445–450

    Article  CAS  PubMed  Google Scholar 

  • Kazi M, Drego L, Nikam C, Ajbani K, Soman R, Shetty A (2015) Molecular characterization of carbapenem-resistant Enterobacteriaceae at a tertiary care laboratory in Mumbai. Eur J Clin Microbiol Infect Dis 34:467–472

    Article  CAS  PubMed  Google Scholar 

  • Miller S, Humphries RM (2016) Clinical laboratory detection of carbapenem-resistant and carbapenemase-producing Enterobacteriaceae. Expert Rev Anti-Infect Ther 14:705–717

    Article  CAS  PubMed  Google Scholar 

  • Pantel A, Souzy D, Sotto A, Lavigne JP (2015) Evaluation of two phenotypic screening tests for carbapenemase-producing Enterobacteriaceae. J Clin Microbiol 53:3359–3362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Patel G, Bonomo RA (2013) “Stormy waters ahead”: global emergence of carbapenemases. Front Microbiol l4:1–17

    Google Scholar 

  • Peter S, Lacher A, Marschal M, Hölzl F, Buhl M, Autenrieth I (2014) Evaluation of phenotypic detection methods for metallo-β-lactamases (MBLs) in clinical isolates of Pseudomonas aeruginosa. Eur J Clin Microbiol Infect Dis 33:1133–1141

    Article  CAS  PubMed  Google Scholar 

  • Poirel L, Walsh TR, Cuvillier V, Nordmann P (2011) Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 70:119–123

    Article  CAS  PubMed  Google Scholar 

  • Ramana KV, Rao R, Sharada CV, Kareem M, Reddy LR, Ratna Mani M (2013) Modified Hodge test: a useful and the low-cost phenotypic method for detection of carbapenemase producers in Enterobacteriaceae members. J Nat Sci Biol Med 4:346–348

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rezaee MA, Pajand O, Nahaei MR, Mahdian R, Aghazadeh M, Ghojazadeh M (2013) Prevalence of ambler class a β-lactamases and ampC expression in cephalosporin-resistant isolates of Acinetobacter baumannii. Diagn Microbiol Infect Dis 76:330–334

    Article  CAS  PubMed  Google Scholar 

  • Shokri D, Rabbani Khorasgani M, Fatemi SM, Soleimani-Delfan A (2017) Resistotyping, phenotyping and genotyping of New Delhi metallo-β-lactamase (NDM) among gram-negative bacilli from Iranian patients. J Med Microbiol 66:402–411

    Article  PubMed  Google Scholar 

  • Singh-Moodley A, Perovic O (2016) Antimicrobial susceptibility testing in predicting the presence of carbapenemase genes in Enterobacteriaceae in South Africa. BMC Infect Dis 16:536–540

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • VanDijk K, Voets GM, Scharringa J, Voskuil S, Fluit AC, Rottier WC (2014) 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 20:345–349

    Article  CAS  Google Scholar 

  • Vlek AL, Frentz D, Haenen A, Bootsma HJ, Notermans DW, Frakking FN (2016) Detection and epidemiology of carbapenemase producing Enterobacteriaceae in the Netherlands in 2013–2014. Eur J Clin Microbiol Infect Dis 35:1089–1096

    Article  CAS  PubMed  Google Scholar 

  • Woodford N, Fagan EJ, Ellington MJ (2006) Multiplex PCR for rapid detection of genes encoding CTX-M extended-spectrum β-lactamases. J Antimicrob Chemother 57:154–155

    Article  CAS  PubMed  Google Scholar 

  • Zong Z, Zhang X (2013) blaNDM-1-carrying Acinetobacter johnsonii detected in hospital sewage. J Antimicrob Chemother 68:1007–1010

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank the “Deputy of Research” of Semnan University of Medical Sciences for their cooperation and for providing facilities to this work.

Funding

This work was supported fully by Semnan University of Medical Sciences (grant nos.730 and 919). This is a report of a database from research projects entitled “Investigation of the prevalence, antimicrobial susceptibility testing and carbapenemase production among Enterobacteriaceae and Pseudomonas aeruginosa isolates recovered from clinical specimens collected from Semnan hospitals and the effect of driving factors” and “Determination of Minimum Inhibitory Concentration (MIC) of Ertapenem in carbapenem non-susceptible isolates of Enterobacteriaceae recovered from hospitalized patients” registered in Semnan University of Medical Sciences, Semnan, Iran.

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Correspondence to Bruno S. Lopes or Omid Pajand.

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The authors declare that they have no conflict of interest.

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The present study was approved by the Semnan University of Medical Sciences, Iran.

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Written informed consent was not necessary for this study. Patient data were anonymous for the purposes of this analysis, and all confidential patients’ information was protected in accordance with the Iranian law.

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Hojabri, Z., Arab, M., Darabi, N. et al. Evaluation of the commercial combined disk test and minimum inhibitory concentration (MIC) determination for detection of carbapenemase producers among Gram-negative bacilli isolated in a region with high prevalence of blaOXA-48 and blaNDM. Int Microbiol 22, 81–89 (2019). https://doi.org/10.1007/s10123-018-0030-1

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  • DOI: https://doi.org/10.1007/s10123-018-0030-1

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