1887

Abstract

, and are clinically relevant members of the (Acb) complex and important nosocomial pathogens. These three species are genetically closely related and phenotypically similar; however, they differ in their epidemiology, antibiotic resistance and pathogenicity. In this study, we investigated the use of a multiplex PCR-based assay designed to detect internal fragments of the 16S–23S rRNA intergenic region and the and genes. The assay was capable of differentiating , and in a reliable manner. In 23 different reference strains and 89 clinical isolates of species, the assay accurately identified clinically relevant Acb complex species except those ‘between 1 and 3’ or ‘close to 13TU’. None of the non-Acb complex species was misidentified. In an analysis of 1034 positive blood cultures, the assay had a sensitivity of 92.4 % and specificity of 98.2 % for Acb complex identification. Our results show that a single multiplex PCR assay can reliably differentiate clinically relevant Acb complex species. Thus, this method may be used to better understand the clinical differences between infections caused by these species.

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2014-09-01
2024-11-04
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References

  1. Chang H. C., Wei Y. F., Dijkshoorn L., Vaneechoutte M., Tang C. T., Chang T. C. 2005; Species-level identification of isolates of the Acinetobacter calcoaceticus–Acinetobacter baumannii complex by sequence analysis of the 16S-23S rRNA gene spacer region. J Clin Microbiol 43:1632–1639 [View Article][PubMed]
    [Google Scholar]
  2. Chen T. L., Siu L. K., Wu R. C., Shaio M. F., Huang L. Y., Fung C. P., Lee C. M., Cho W. L. 2007; Comparison of one-tube multiplex PCR, automated ribotyping and intergenic spacer (ITS) sequencing for rapid identification of Acinetobacter baumannii. Clin Microbiol Infect 13:801–806 [View Article][PubMed]
    [Google Scholar]
  3. Chen T. L., Lee Y. T., Kuo S. C., Hsueh P. R., Chang F. Y., Siu L. K., Ko W. C., Fung C. P. 2010; Emergence and distribution of plasmids bearing the blaOXA-51-like gene with an upstream ISAba1 in carbapenem-resistant Acinetobacter baumannii isolates in Taiwan. Antimicrob Agents Chemother 54:4575–4581 [View Article][PubMed]
    [Google Scholar]
  4. Chuang Y. C., Sheng W. H., Li S. Y., Lin Y. C., Wang J. T., Chen Y. C., Chang S. C. 2011; Influence of genospecies of Acinetobacter baumannii complex on clinical outcomes of patients with Acinetobacter bacteremia. Clin Infect Dis 52:352–360 [View Article][PubMed]
    [Google Scholar]
  5. Dijkshoorn L., Van Harsselaar B., Tjernberg I., Bouvet P. J., Vaneechoutte M. 1998; Evaluation of amplified ribosomal DNA restriction analysis for identification of Acinetobacter genomic species. Syst Appl Microbiol 21:33–39 [View Article][PubMed]
    [Google Scholar]
  6. Dijkshoorn L., Nemec A., Seifert H. 2007; An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nat Rev Microbiol 5:939–951 [View Article][PubMed]
    [Google Scholar]
  7. Fredricks D. N., Relman D. A. 1998; Improved amplification of microbial DNA from blood cultures by removal of the PCR inhibitor sodium polyanetholesulfonate. J Clin Microbiol 36:2810–2816[PubMed]
    [Google Scholar]
  8. Gerner-Smidt P., Tjernberg I., Ursing J. 1991; Reliability of phenotypic tests for identification of Acinetobacter species. J Clin Microbiol 29:277–282[PubMed]
    [Google Scholar]
  9. Higgins P. G., Wisplinghoff H., Krut O., Seifert H. 2007; A PCR-based method to differentiate between Acinetobacter baumannii and Acinetobacter genomic species 13TU. Clin Microbiol Infect 13:1199–1201 [View Article][PubMed]
    [Google Scholar]
  10. Higgins P. G., Lehmann M., Wisplinghoff H., Seifert H. 2010; gyrB multiplex PCR to differentiate between Acinetobacter calcoaceticus and Acinetobacter genomic species 3. J Clin Microbiol 48:4592–4594 [View Article][PubMed]
    [Google Scholar]
  11. Huang L. Y., Chen T. L., Lu P. L., Tsai C. A., Cho W. L., Chang F. Y., Fung C. P., Siu L. K. 2008; Dissemination of multidrug-resistant, class 1 integron-carrying Acinetobacter baumannii isolates in Taiwan. Clin Microbiol Infect 14:1010–1019 [View Article][PubMed]
    [Google Scholar]
  12. Krawczyk B., Lewandowski K., Kur J. 2002; Comparative studies of the Acinetobacter genus and the species identification method based on the recA sequences. Mol Cell Probes 16:1–11 [View Article][PubMed]
    [Google Scholar]
  13. Kuo S. C., Lee Y. T., Yang S. P., Chiang M. C., Lin Y. T., Tseng F. C., Chen T. L., Fung C. P. 2013; Evaluation of the effect of appropriate antimicrobial therapy on mortality associated with Acinetobacter nosocomialis bacteraemia. Clin Microbiol Infect 19:634–639 [View Article][PubMed]
    [Google Scholar]
  14. Lee N. Y., Chang T. C., Wu C. J., Chang C. M., Lee H. C., Chen P. L., Lee C. C., Ko N. Y., Ko W. C. 2010; Clinical manifestations, antimicrobial therapy, and prognostic factors of monomicrobial Acinetobacter baumannii complex bacteremia. J Infect 61:219–227 [View Article][PubMed]
    [Google Scholar]
  15. Lee Y.-T., Kuo S.-C., Chiang M.-C., Yang S.-P., Chen C.-P., Chen T.-L., Fung C.-P. 2012a; Emergence of carbapenem-resistant non-baumannii species of Acinetobacter harboring a blaOXA-51-like gene that is intrinsic to A. baumannii. Antimicrob Agents Chemother 56:1124–1127 [View Article][PubMed]
    [Google Scholar]
  16. Lee Y. T., Kuo S. C., Yang S. P., Lin Y. T., Tseng F. C., Chen T. L., Fung C. P. 2012b; Impact of appropriate antimicrobial therapy on mortality associated with Acinetobacter baumannii bacteremia: relation to severity of infection. Clin Infect Dis 55:209–215 [View Article][PubMed]
    [Google Scholar]
  17. Nemec A., Dijkshoorn L., Cleenwerck I., De Baere T., Janssens D., Van Der Reijden T. J., Jezek P., Vaneechoutte M. 2003; Acinetobacter parvus sp. nov., a small-colony-forming species isolated from human clinical specimens. Int J Syst Evol Microbiol 53:1563–1567 [View Article][PubMed]
    [Google Scholar]
  18. Nemec A., Musílek M., Maixnerová M., De Baere T., van der Reijden T. J., Vaneechoutte M., Dijkshoorn L. 2009; Acinetobacter beijerinckii sp. nov. and Acinetobacter gyllenbergii sp. nov., haemolytic organisms isolated from humans. Int J Syst Evol Microbiol 59:118–124 [View Article][PubMed]
    [Google Scholar]
  19. Nemec A., Musílek M., Sedo O., De Baere T., Maixnerová M., van der Reijden T. J., Zdráhal Z., Vaneechoutte M., Dijkshoorn L. 2010; Acinetobacter bereziniae sp. nov. and Acinetobacter guillouiae sp. nov., to accommodate Acinetobacter genomic species 10 and 11, respectively. Int J Syst Evol Microbiol 60:896–903 [View Article][PubMed]
    [Google Scholar]
  20. Nemec A., Krizova L., Maixnerova M., van der Reijden T. J., Deschaght P., Passet V., Vaneechoutte M., Brisse S., Dijkshoorn L. 2011; Genotypic and phenotypic characterization of the Acinetobacter calcoaceticusAcinetobacter baumannii complex with the proposal of Acinetobacter pittii sp. nov. (formerly Acinetobacter genomic species 3) and Acinetobacter nosocomialis sp. nov. (formerly Acinetobacter genomic species 13TU). Res Microbiol 162:393–404 [View Article][PubMed]
    [Google Scholar]
  21. Peleg A. Y., Seifert H., Paterson D. L. 2008; Acinetobacter baumannii: emergence of a successful pathogen. Clin Microbiol Rev 21:538–582 [View Article][PubMed]
    [Google Scholar]
  22. Šedo O., Nemec A., Křížová L., Kačalová M., Zdráhal Z. 2013; Improvement of MALDI-TOF MS profiling for the differentiation of species within the Acinetobacter calcoaceticusAcinetobacter baumannii complex. Syst Appl Microbiol 36:572–578 [View Article][PubMed]
    [Google Scholar]
  23. Turton J. F., Woodford N., Glover J., Yarde S., Kaufmann M. E., Pitt T. L. 2006; Identification of Acinetobacter baumannii by detection of the blaOXA-51-like carbapenemase gene intrinsic to this species. J Clin Microbiol 44:2974–2976 [View Article][PubMed]
    [Google Scholar]
  24. Vaneechoutte M., Dijkshoorn L., Tjernberg I., Elaichouni A., de Vos P., Claeys G., Verschraegen G. 1995; Identification of Acinetobacter genomic species by amplified ribosomal DNA restriction analysis. J Clin Microbiol 33:11–15[PubMed]
    [Google Scholar]
  25. Wisplinghoff H., Paulus T., Lugenheim M., Stefanik D., Higgins P. G., Edmond M. B., Wenzel R. P., Seifert H. 2012; Nosocomial bloodstream infections due to Acinetobacter baumannii, Acinetobacter pittii and Acinetobacter nosocomialis in the United States. J Infect 64:282–290 [View Article][PubMed]
    [Google Scholar]
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