1887

Abstract

To understand the diversity of porin disruption in , the major outer membrane protein (OMP) porins, OmpK35 and OmpK36, were examined in a set of isolates that did not harbour traditional carbapenem-hydrolysing enzymes, but nevertheless tested non-susceptible to ertapenem.

A world-wide collection of isolates that were part of the Study for Monitoring Antimicrobial Resistance Trends (SMART) surveillance project over the years 2008–2014 were characterised with regard to their β-lactamase gene carriage and potential permeability defects. Four hundred and eighty-seven isolates that did not carry carbapenemase genes, but were non-susceptible to ertapenem, were investigated by sequence analysis of the genes encoding OmpK35 and OmpK36. Isolates without obvious genetic lesions in either major porin gene were further examined by outer membrane protein SDS-PAGE.

The majority of isolates, 83.0 % (404/487), exhibited clear genetic disruption in either or both of the and genes. Among the proportion of the collection with the highest ertapenem MIC value (>4 mg l), 60.5 % (115/190) showed mutation in both porin genes. Isolates without obvious genetic mutations were examined by SDS-PAGE, and 90.4 % (75/83) were found to lack or show altered expression of at least one of the major OMPs when compared to an ertapenem sensitive control strain.

This study illustrates that porin deficiency in is a widespread phenomenon, and in combination with ESBLs and/or AmpC enzymes, likely accounts for the elevated ertapenem MICs observed in this study.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000691
2018-03-01
2024-12-02
Loading full text...

Full text loading...

/deliver/fulltext/jmm/67/3/289.html?itemId=/content/journal/jmm/10.1099/jmm.0.000691&mimeType=html&fmt=ahah

References

  1. Bradford PA, Urban C, Mariano N, Projan SJ, Rahal JJ et al. Imipenem resistance in Klebsiella pneumoniae is associated with the combination of ACT-1, a plasmid-mediated AmpC beta-lactamase, and the foss of an outer membrane protein. Antimicrob Agents Chemother 1997; 41:563–569[PubMed]
    [Google Scholar]
  2. Cao VT, Arlet G, Ericsson BM, Tammelin A, Courvalin P et al. Emergence of imipenem resistance in Klebsiella pneumoniae owing to combination of plasmid-mediated CMY-4 and permeability alteration. J Antimicrob Chemother 2000; 46:895–900 [View Article][PubMed]
    [Google Scholar]
  3. Jacoby GA, Mills DM, Chow N. Role of β-lactamases and porins in resistance to ertapenem and other beta-lactams in Klebsiella pneumoniae. Antimicrob Agents Chemother 2004; 48:3203–3206 [View Article][PubMed]
    [Google Scholar]
  4. Matsumura Y, Tanaka M, Yamamoto M, Nagao M, Machida K et al. High prevalence of carbapenem resistance among plasmid-mediated AmpC β-lactamase-producing Klebsiella pneumoniae during outbreaks in liver transplantation units. Int J Antimicrob Agents 2015; 45:33–40 [View Article][PubMed]
    [Google Scholar]
  5. Martínez-Martínez L, Pascual A, Hernández-Allés S, Alvarez-Díaz D, Suárez AI et al. Roles of β-lactamases and porins in activities of carbapenems and cephalosporins against Klebsiella pneumoniae. Antimicrob Agents Chemother 1999; 43:1669–1673[PubMed]
    [Google Scholar]
  6. Doumith M, Ellington MJ, Livermore DM, Woodford N. Molecular mechanisms disrupting porin expression in ertapenem-resistant Klebsiella and Enterobacter spp. clinical isolates from the UK. J Antimicrob Chemother 2009; 63:659–667 [View Article][PubMed]
    [Google Scholar]
  7. Landman D, Bratu S, Quale J. Contribution of OmpK36 to carbapenem susceptibility in KPC-producing Klebsiella pneumoniae. J Med Microbiol 2009; 58:1303–1308 [View Article][PubMed]
    [Google Scholar]
  8. Elliott E, Brink AJ, van Greune J, Els Z, Woodford N et al. In vivo development of ertapenem resistance in a patient with pneumonia caused by Klebsiella pneumoniae with an extended-spectrum β-lactamase. Clin Infect Diseases 2006; 42:e95-e98 [View Article]
    [Google Scholar]
  9. Livermore DM. Current epidemiology and growing resistance of gram-negative pathogens. Korean J Intern Med 2012; 27:128–142 [View Article][PubMed]
    [Google Scholar]
  10. García-Fernández A, Miriagou V, Papagiannitsis CC, Giordano A, Venditti M et al. An ertapenem-resistant extended-spectrum-β-lactamase-producing Klebsiella pneumoniae clone carries a novel OmpK36 porin variant. Antimicrob Agents Chemother 2010; 54:4178–4184 [View Article][PubMed]
    [Google Scholar]
  11. Novais A, Rodrigues C, Branquinho R, Antunes P, Grosso F et al. Spread of an OmpK36-modified ST15 Klebsiella pneumoniae variant during an outbreak involving multiple carbapenem-resistant Enterobacteriaceae species and clones. Eur J Clin Microbiol Infect Dis 2012; 31:3057–3063 [View Article][PubMed]
    [Google Scholar]
  12. Poulou A, Voulgari E, Vrioni G, Koumaki V, Xidopoulos G et al. Outbreak caused by an ertapenem-resistant, CTX-M-15-producing Klebsiella pneumoniae sequence type 101 clone carrying an OmpK36 porin variant. J Clin Microbiol 2013; 51:3176–3182 [View Article][PubMed]
    [Google Scholar]
  13. Morrissey I, Hackel M, Badal R, Bouchillon S, Hawser S et al. A review of ten years of the study for monitoring antimicrobial resistance trends (SMART) from 2002 to 2011. Pharmaceuticals 2013; 6:1335–1346 [View Article][PubMed]
    [Google Scholar]
  14. Lob SH, Hackel MA, Kazmierczak KM, Young K, Motyl MR et al. In vitro activity of imipenem-relebactam against gram-negative ESKAPE pathogens isolated by clinical laboratories in the United States in 2015 (results from the SMART global surveillance program). Antimicrob Agents Chemother 2017; 61:e02209-16 [View Article][PubMed]
    [Google Scholar]
  15. Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Susceptibility Testing, twenty-seventh informational supplement. CLSI document M100-S27 Wayne, PA: Clinical and Laboratory Standards Institute; 2017
    [Google Scholar]
  16. Dallenne C, Da Costa A, Decré D, Favier C, Arlet G. Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J Antimicrob Chemother 2010; 65:490–495 [View Article][PubMed]
    [Google Scholar]
  17. Ellington MJ, Kistler J, Livermore DM, Woodford N. Multiplex PCR for rapid detection of genes encoding acquired metallo-β-lactamases. J Antimicrob Chemother 2007; 59:321–322 [View Article][PubMed]
    [Google Scholar]
  18. Kazmierczak KM, Lob SH, Hoban DJ, Hackel MA, Badal RE et al. Characterization of extended-spectrum β-lactamases and antimicrobial resistance of Klebsiella pneumoniae in intra-abdominal infection isolates in Latin America, 2008–2012. Results of the study for monitoring antimicrobial resistance trends. Diagn Microbiol Infect Dis 2015; 82:209–214 [View Article][PubMed]
    [Google Scholar]
  19. Lascols C, Hackel M, Marshall SH, Hujer AM, Bouchillon S et al. Increasing prevalence and dissemination of NDM-1 metallo-β-lactamase in India: data from the SMART study (2009). J Antimicrob Chemother 2011; 66:1992–1997 [View Article][PubMed]
    [Google Scholar]
  20. Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 2002; 40:2153–2162 [View Article][PubMed]
    [Google Scholar]
  21. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011; 70:119–123 [View Article][PubMed]
    [Google Scholar]
  22. Woodford N, Fagan EJ, Ellington MJ. Multiplex PCR for rapid detection of genes encoding CTX-M extended-spectrum β-lactamases. J Antimicrob Chemother 2006; 57:154–155 [View Article][PubMed]
    [Google Scholar]
  23. Yigit H, Queenan AM, Anderson GJ, Domenech-Sanchez A, Biddle JW et al. Novel carbapenem-hydrolyzing β-lactamase, KPC-1, from a carbapenem-resistant strain of Klebsiella pneumoniae. Antimicrob Agents Chemother 2001; 45:1151–1161 [View Article][PubMed]
    [Google Scholar]
  24. Clancy CJ, Chen L, Hong JH, Cheng S, Hao B et al. Mutations of the ompK36 porin gene and promoter impact responses of sequence type 258, KPC-2-producing Klebsiella pneumoniae strains to doripenem and doripenem-colistin. Antimicrob Agents Chemother 2013; 57:5258–5265 [View Article][PubMed]
    [Google Scholar]
  25. Carlone GM, Thomas ML, Rumschlag HS, Sottnek FO. Rapid microprocedure for isolating detergent-insoluble outer membrane proteins from Haemophilus species. J Clin Microbiol 1986; 24:330–332[PubMed]
    [Google Scholar]
  26. Woodford N, Tierno PM, Young K, Tysall L, Palepou MF et al. Outbreak of Klebsiella pneumoniae producing a new carbapenem-hydrolyzing class A β-lactamase, KPC-3, in a New York medical center. Antimicrob Agents Chemother 2004; 48:4793–4799 [View Article][PubMed]
    [Google Scholar]
  27. Hernández-Allés S, Albertí S, Alvarez D, Doménech-Sánchez A, Martínez-Martínez L et al. Porin expression in clinical isolates of Klebsiella pneumoniae. Microbiology 1999; 145:673–679 [View Article][PubMed]
    [Google Scholar]
  28. Tsai YK, Fung CP, Lin JC, Chen JH, Chang FY et al. Klebsiella pneumoniae outer membrane porins OmpK35 and OmpK36 play roles in both antimicrobial resistance and virulence. Antimicrob Agents Chemother 2011; 55:1485–1493 [View Article][PubMed]
    [Google Scholar]
  29. Zhang Y, Jiang X, Wang Y, Li G, Tian Y et al. Contribution of β-lactamases and porin proteins OmpK35 and OmpK36 to carbapenem resistance in clinical isolates of KPC-2-producing Klebsiella pneumoniae. Antimicrob Agents Chemother 2014; 58:1214–1217 [View Article][PubMed]
    [Google Scholar]
  30. Hernández-Allés S, Conejo M, Pascual A, Tomás JM, Benedí VJ et al. Relationship between outer membrane alterations and susceptibility to antimicrobial agents in isogenic strains of Klebsiella pneumoniae. J Antimicrob Chemother 2000; 46:273–277 [View Article][PubMed]
    [Google Scholar]
  31. Wang XD, Cai JC, Zhou HW, Zhang R, Chen GX. Reduced susceptibility to carbapenems in Klebsiella pneumoniae clinical isolates associated with plasmid-mediated β -lactamase production and OmpK36 porin deficiency. J Med Microbiol 2009; 58:1196–1202 [View Article][PubMed]
    [Google Scholar]
  32. Leavitt A, Chmelnitsky I, Colodner R, Ofek I, Carmeli Y et al. Ertapenem resistance among extended-spectrum-β-lactamase-producing Klebsiella pneumoniae isolates. J Clin Microbiol 2009; 47:969–974 [View Article][PubMed]
    [Google Scholar]
  33. Sho T, Muratani T, Hamasuna R, Yakushiji H, Fujimoto N et al. The mechanism of high-level carbapenem resistance in Klebsiella pneumoniae: underlying Ompk36-deficient strains represent a threat of emerging high-level carbapenem-resistant K. pneumoniae with IMP-1 β-lactamase production in Japan. Microb Drug Resist 2013; 19:274–281 [View Article][PubMed]
    [Google Scholar]
  34. Yang D, Guo Y, Zhang Z. Combined porin loss and extended spectrum β-lactamase production is associated with an increasing imipenem minimal inhibitory concentration in clinical Klebsiella pneumoniae strains. Curr Microbiol 2009; 58:366–370 [View Article][PubMed]
    [Google Scholar]
  35. Tsai YK, Liou CH, Fung CP, Lin JC, Siu LK. Single or in combination antimicrobial resistance mechanisms of Klebsiella pneumoniae contribute to varied susceptibility to different carbapenems. PLoS One 2013; 8:e79640 [View Article][PubMed]
    [Google Scholar]
  36. Mena A, Plasencia V, García L, Hidalgo O, Ayestarán JI et al. Characterization of a large outbreak by CTX-M-1-producing Klebsiella pneumoniae and mechanisms leading to in vivo carbapenem resistance development. J Clin Microbiol 2006; 44:2831–2837 [View Article][PubMed]
    [Google Scholar]
  37. Agyekum A, Fajardo-Lubián A, Ai X, Ginn AN, Zong Z et al. Predictability of phenotype in relation to common β-lactam resistance mechanisms in Escherichia coli and Klebsiella pneumoniae. J Clin Microbiol 2016; 54:1243–1250 [View Article][PubMed]
    [Google Scholar]
  38. Doménech-Sánchez A, Martínez-Martínez L, Hernández-Allés S, del Carmen Conejo M, Pascual A et al. Role of Klebsiella pneumoniae OmpK35 porin in antimicrobial resistance. Antimicrob Agents Chemother 2003; 47:3332–3335 [View Article][PubMed]
    [Google Scholar]
  39. Albertí S, Rodríquez-Quiñones F, Schirmer T, Rummel G, Tomás JM et al. A porin from Klebsiella pneumoniae: sequence homology, three-dimensional model, and complement binding. Infect Immun 1995; 63:903–910[PubMed]
    [Google Scholar]
  40. Dutzler R, Rummel G, Albertí S, Hernández-Allés S, Phale P et al. Crystal structure and functional characterization of OmpK36, the osmoporin of Klebsiella pneumoniae. Structure 1999; 7:425–434 [View Article][PubMed]
    [Google Scholar]
  41. Bredin J, Saint N, Malléa M, de E, Molle G et al. Alteration of pore properties of Escherichia coli OmpF induced by mutation of key residues in anti-loop 3 region. Biochem J 2002; 363:521–528 [View Article][PubMed]
    [Google Scholar]
  42. E, Baslé A, Jaquinod M, Saint N, Malléa M et al. A new mechanism of antibiotic resistance in Enterobacteriaceae induced by a structural modification of the major porin. Mol Microbiol 2001; 41:189–198 [View Article][PubMed]
    [Google Scholar]
  43. Thiolas A, Bornet C, Davin-Régli A, Pagès JM, Bollet C. Resistance to imipenem, cefepime, and cefpirome associated with mutation in Omp36 osmoporin of Enterobacter aerogenes. Biochem Biophys Res Commun 2004; 317:851–856 [View Article][PubMed]
    [Google Scholar]
  44. Filgona J, Banerjee T, Anupurba S. Role of efflux pumps inhibitor in decreasing antibiotic resistance of Klebsiella pneumoniae in a tertiary hospital in North India. J Infect Dev Ctries 2015; 9:815–820 [View Article][PubMed]
    [Google Scholar]
  45. Smith SG, Mahon V, Lambert MA, Fagan RP. A molecular Swiss army knife: OmpA structure, function and expression. FEMS Microbiol Lett 2007; 273:1–11 [View Article][PubMed]
    [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.000691
Loading
/content/journal/jmm/10.1099/jmm.0.000691
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error