1887

Abstract

A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D β-lactamases ( -like, -like, -like and -like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant isolates. Well-characterized strains of were used as positive controls and non- strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMP-resistant isolates were positive for the -like gene, and one isolate (2 %) was positive for the -like gene. The -like gene was not detected in any of the 46 IMP-resistant strains and the -like gene was identified in both IMP-resistant and non-resistant . All 11 non- bacteria produced a negative result for each of the four genes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in and potentially other clinically significant multidrug-resistant Gram-negative bacteria.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.045823-0
2012-11-01
2020-11-30
Loading full text...

Full text loading...

/deliver/fulltext/jmm/61/11/1532.html?itemId=/content/journal/jmm/10.1099/jmm.0.045823-0&mimeType=html&fmt=ahah

References

  1. Abbo A., Navon-Venezia S., Hammer-Muntz O., Krichali T., Siegman-Igra Y., Carmeli Y. 2005; Multidrug-resistant Acinetobacter baumannii. Emerg Infect Dis 11:22–29 [CrossRef][PubMed]
    [Google Scholar]
  2. Adams M. D., Goglin K., Molyneaux N., Hujer K. M., Lavender H., Jamison J. J., MacDonald I. J., Martin K. M., Russo T. other authors 2008; Comparative genome sequence analysis of multidrug-resistant Acinetobacter baumannii. J Bacteriol 190:8053–8064 [CrossRef][PubMed]
    [Google Scholar]
  3. Barraud O., Baclet M. C., Denis F., Ploy M. C. 2010; Quantitative multiplex real-time PCR for detecting class 1, 2 and 3 integrons. J Antimicrob Chemother 65:1642–1645 [CrossRef][PubMed]
    [Google Scholar]
  4. Bush K., Jacoby G. A. 2010; Updated functional classification of β-lactamases. Antimicrob Agents Chemother 54:969–976 [CrossRef][PubMed]
    [Google Scholar]
  5. Davis K. A., Moran K. A., McAllister C. K., Gray P. J. 2005; Multidrug-resistant Acinetobacter extremity infections in soldiers. Emerg Infect Dis 11:1218–1224 [CrossRef][PubMed]
    [Google Scholar]
  6. Fournier P. E., Vallenet D., Barbe V., Audic S., Ogata H., Poirel L., Richet H., Robert C., Mangenot S. other authors 2006; Comparative genomics of multidrug resistance in Acinetobacter baumannii. PLoS Genet 2:e7 [CrossRef][PubMed]
    [Google Scholar]
  7. Frye J. G., Lindsey R. L., Rondeau G., Porwollik S., Long F., McClelland M., Jackson C. R., Englen M. D., Meinersmann R. J. other authors 2010; Development of a DNA microarray to detect antimicrobial resistance genes identified in the National Center for Biotechnology Information database. Microb Drug Resist 16:9–19 [CrossRef][PubMed]
    [Google Scholar]
  8. Higgins P. G., Dammhayn C., Hackel M., Seifert H. 2010; Global spread of carbapenem-resistant Acinetobacter baumannii. J Antimicrob Chemother 65:233–238 [CrossRef][PubMed]
    [Google Scholar]
  9. Huang X.-Z., Frye J. G., Chahine M. A., Cash D. M., Barber M. G., Babel B. S., Kasper M. R., Whitman T. J., Lindler L. E. other authors 2010; Genotypic and phenotypic correlations of multidrug-resistant Acinetobacter baumannii-A. calcoaceticus complex strains isolated from patients at the National Naval Medical Center. J Clin Microbiol 48:4333–4336 [CrossRef][PubMed]
    [Google Scholar]
  10. Huang X.-Z., Chahine M. A., Frye J. G., Cash D. M., Lesho E. P., Craft D. W., Lindler L. E., Nikolich M. P. 2012; Molecular analysis of imipenem-resistant Acinetobacter baumannii isolated from US service members wounded in Iraq, 2003–2008. Epidemiol Infect1–6 [CrossRef][PubMed]
    [Google Scholar]
  11. Iacono M., Villa L., Fortini D., Bordoni R., Imperi F., Bonnal R. J., Sicheritz-Ponten T., De Bellis G., Visca P. other authors 2008; Whole-genome pyrosequencing of an epidemic multidrug-resistant Acinetobacter baumannii strain belonging to the European clone II group. Antimicrob Agents Chemother 52:2616–2625 [CrossRef][PubMed]
    [Google Scholar]
  12. Mostachio A. K., van der Heidjen I., Rossi F., Levin A. S., Costa S. F. 2009; Multiplex PCR for rapid detection of genes encoding oxacillinases and metallo-β-lactamases in carbapenem-resistant Acinetobacter spp. J Med Microbiol 58:1522–1524 [CrossRef][PubMed]
    [Google Scholar]
  13. Poirel L., Nordmann P. 2006; Carbapenem resistance in Acinetobacter baumannii: mechanisms and epidemiology. Clin Microbiol Infect 12:826–836 [CrossRef][PubMed]
    [Google Scholar]
  14. Poirel L., Naas T., Nordmann P. 2010; Class D β-lactamases: diversity, epidemiology and genetics. Antimicrob Agents Chemother 54:24–38 [CrossRef][PubMed]
    [Google Scholar]
  15. Richet H., Fournier P. E. 2006; Nosocomial infections caused by Acinetobacter baumannii: a major threat worldwide. Infect Control Hosp Epidemiol 27:645–646 [CrossRef][PubMed]
    [Google Scholar]
  16. Scott P., Deye G., Srinivasan A., Murray C., Moran K., Hulten E., Fishbain J., Craft D., Riddell S. other authors 2007; An outbreak of multidrug-resistant Acinetobacter baumannii–calcoaceticus complex infection in the US military health care system associated with military operations in Iraq. Clin Infect Dis 44:1577–1584 [CrossRef][PubMed]
    [Google Scholar]
  17. Tenover F. C., Kalsi R. K., Williams P. P., Carey R. B., Stocker S., Lonsway D., Rasheed J. K., Biddle J. W., McGowan J. E. Jr, Hanna B. 2006; Carbapenem resistance in Klebsiella pneumoniae not detected by automated susceptibility testing. Emerg Infect Dis 12:1209–1213 [CrossRef][PubMed]
    [Google Scholar]
  18. Turton J. F., Ward M. E., Woodford N., Kaufmann M. E., Pike R., Livermore D. M., Pitt T. L. 2006a; The role of ISAba1 in expression of OXA carbapenemase genes in Acinetobacter baumannii. FEMS Microbiol Lett 258:72–77 [CrossRef][PubMed]
    [Google Scholar]
  19. Turton J. F., Woodford N., Glover J., Yarde S., Kaufmann M. E., Pitt T. L. 2006b; Identification of Acinetobacter baumannii by detection of the blaOXA-51-like carbapenemase gene intrinsic to this species. J Clin Microbiol 44:2974–2976 [CrossRef][PubMed]
    [Google Scholar]
  20. Woodford N., Ellington M. J., Coelho J. M., Turton J. F., Ward M. E., Brown S., Amyes S. G., Livermore D. M. 2006; Multiplex PCR for genes encoding prevalent OXA carbapenemases in Acinetobacter spp. Int J Antimicrob Agents 27:351–353 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.045823-0
Loading
/content/journal/jmm/10.1099/jmm.0.045823-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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