Defining criteria to interpret multilocus variable-number tandem repeat analysis to aid outbreak investigation Free

Abstract

PFGE is currently the North American standard for surveillance for but lacks discriminatory power to aid outbreak investigation. A further limitation to PFGE is the high baseline rate of the epidemic North American pulsotype (NAP) 1 strain in hospitals. Multilocus variable-number tandem repeat analysis (MLVA) appears to have superior discriminatory power but criteria to define clonality have not been set. We conducted surveillance for toxin-positive infection (CDI) at a single academic health sciences centre between September 2009 and April 2010. Seventy-four patient specimens resulting in 86 discrete CDI episodes were subjected to PFGE and MLVA. Results were analysed using Bionumerics software to generate phylogenetic trees and coupled to patient demographic data. Amongst the NAP1 strains, two distinct clusters were identified by MLVA using 90 % similarity as a cut-off by Manhattan distance-based clustering, four clusters using 95 % and seven clusters using 97 %. Population analysis conducted on multiple colonies ( = 25) demonstrated that 1–3 % difference in MLVA types was typical for a single individual. Typing was also conducted in the context of institutional outbreaks ( = 42, three outbreaks) in order to determine clusters within the NAP1 strain. By combining longitudinal surveillance with epidemiological information, single specimen population analysis and typing in the context of institutional outbreaks, we conclude that the use of the Manhattan distance-based clustering with a cut-off of 95–97 % is capable of distinguishing outbreak clones from sporadic isolates.

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2011-08-01
2024-03-28
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References

  1. Antikainen J., Pasanen T., Mero S., Tarkka E., Kirveskari J., Kotila S., Mentula S., Könönen E., Virolainen-Julkunen A. R. et al. 2009; Detection of virulence genes of Clostridium difficile by multiplex PCR. APMIS 117:607–613 [View Article][PubMed]
    [Google Scholar]
  2. Aspinall S. T., Hutchinson D. N. 1992; New selective medium for isolating Clostridium difficile from faeces. J Clin Pathol 45:812–814 [View Article][PubMed]
    [Google Scholar]
  3. Broukhanski G., Low D. E., Pillai D. R. 2011; Modified multiple-locus variable-number tandem-repeat analysis for rapid identification and typing of Clostridium difficile during institutional outbreaks. J Clin Microbiol 49:1983–1986 [CrossRef]
    [Google Scholar]
  4. Fawley W. N., Freeman J., Smith C., Harmanus C., van den Berg R. J., Kuijper E. J., Wilcox M. H. 2008; Use of highly discriminatory fingerprinting to analyze clusters of Clostridium difficile infection cases due to epidemic ribotype 027 strains. J Clin Microbiol 46:954–960 [View Article][PubMed]
    [Google Scholar]
  5. Goorhuis A., Legaria M. C., van den Berg R. J., Harmanus C., Klaassen C. H., Brazier J. S., Lumelsky G., Kuijper E. J. 2009; Application of multiple-locus variable-number tandem-repeat analysis to determine clonal spread of toxin A-negative Clostridium difficile in a general hospital in Buenos Aires, Argentina. Clin Microbiol Infect 15:1080–1086 [CrossRef]
    [Google Scholar]
  6. Ichikawa K., Yagi T., Inagaki T., Moriyama M., Nakagawa T., Uchiya K., Nikai T., Ogawa K. 2010; Molecular typing of Mycobacterium intracellulare using multilocus variable-number of tandem-repeat analysis: identification of loci and analysis of clinical isolates. Microbiology 156:496–504 [View Article][PubMed]
    [Google Scholar]
  7. Killgore G., Thompson A., Johnson S., Brazier J., Kuijper E., Pepin J., Frost E. H., Savelkoul P., Nicholson B. et al. 2008; Comparison of seven techniques for typing international epidemic strains of Clostridium difficile: restriction endonuclease analysis, pulsed-field gel electrophoresis, PCR-ribotyping, multilocus sequence typing, multilocus variable-number tandem-repeat analysis, amplified fragment length polymorphism, and surface layer protein A gene sequence typing. J Clin Microbiol 46:431–437 [View Article][PubMed]
    [Google Scholar]
  8. Kuijper E. J., van den Berg R. J., Brazier J. S. 2009; Comparison of molecular typing methods applied to Clostridium difficile . Methods Mol Biol 551:159–171 [View Article][PubMed]
    [Google Scholar]
  9. Marsh J. W., O'Leary M. M., Shutt K. A., Pasculle A. W., Johnson S., Gerding D. N., Muto C. A., Harrison L. H. 2006; Multilocus variable-number tandem-repeat analysis for investigation of Clostridium difficile transmission in hospitals. J Clin Microbiol 44:2558–2566 [CrossRef]
    [Google Scholar]
  10. Mulvey M. R., Boyd D. A., Gravel D., Hutchinson J., Kelly S., McGeer A., Moore D., Simor A., Suh K. N. et al. 2010; Hypervirulent Clostridium difficile strains in hospitalized patients, Canada. Emerg Infect Dis 16:678–681[PubMed] [CrossRef]
    [Google Scholar]
  11. Pillai D. R., Longtin J., Low D. E. 2010; Surveillance data on outbreaks of Clostridium difficile infection in Ontario, Canada, in 2008-2009. Clin Infect Dis 50:1685–1686, author reply 1686 [View Article][PubMed]
    [Google Scholar]
  12. Scaria J., Ponnala L., Janvilisri T., Yan W., Mueller L. A., Chang Y. F. 2010; Analysis of ultra low genome conservation in Clostridium difficile . PLoS ONE 5:e15147 [View Article][PubMed]
    [Google Scholar]
  13. Sebaihia M., Wren B. W., Mullany P., Fairweather N. F., Minton N., Stabler R., Thomson N. R., Roberts A. P., Cerdeño-Tárraga A. M. et al. 2006; The multidrug-resistant human pathogen Clostridium difficile has a highly mobile, mosaic genome. Nat Genet 38:779–786 [View Article][PubMed]
    [Google Scholar]
  14. Tanner H. E., Hardy K. J., Hawkey P. M. 2010; Coexistence of multiple multilocus variable-number tandem-repeat analysis subtypes of Clostridium difficile PCR ribotype 027 strains within fecal specimens. J Clin Microbiol 48:985–987 [View Article][PubMed]
    [Google Scholar]
  15. van den Berg R. J., Schaap I., Templeton K. E., Klaassen C. H., Kuijper E. J. 2007; Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat analysis. J Clin Microbiol 45:1024–1028 [View Article][PubMed]
    [Google Scholar]
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