J Med Microbiol Email Content Delivery
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Slack, A.
Right arrow Articles by Smythe, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Slack, A.
Right arrow Articles by Smythe, L.
Agricola
Right arrow Articles by Slack, A.
Right arrow Articles by Smythe, L.
J Med Microbiol 55 (2006), 1549-1557; DOI: 10.1099/jmm.0.46779-0
© 2006 Society for General Microbiology
ISSN 1473-5644

An improved multiple-locus variable number of tandem repeats analysis for Leptospira interrogans serovar Australis: a comparison with fluorescent amplified fragment length polymorphism analysis and its use to redefine the molecular epidemiology of this serovar in Queensland, Australia

Andrew Slack, Meegan Symonds, Michael Dohnt and Lee Smythe

WHO/FAO/OIE Collaborating Centre for Reference and Research on Leptospirosis, Western Pacific Region, Centre for Public Health Sciences, Queensland Health Scientific Services, Brisbane, Australia

Correspondence
Andrew Slack
andrew_slack{at}health.qld.gov.au

Received 13 June 2006
Accepted 27 July 2006

In this study, an improved multiple-locus variable number of tandem repeats analysis (MLVA) method based upon a previously published method is described. Improvements to the method included redesigned primers and PCR conditions, combined with pooled capillary electrophoresis using multicolored dyes. Allele sizes were converted into an allele string, and each unique allele string was assigned a numerical MLVA type (MVT). The improved MLVA method was then applied to 96 previously characterized Leptospira interrogans serovar Australis isolates from human and animal sources. The improved MLVA was found to have between six and 13 alleles at each locus, compared with three to eight in the original. The mean Hunter–Gaston diversity index (HGDI) for the improved MLVA method was 0.654, compared with 0.599 in the original; this increase in diversity was largely due to changes in the analysis of the variable number of tandem repeat (VNTR) data. When the improved MLVA method was compared with the fluorescent amplified fragment length polymorphism (FAFLP) method, there was a high level of concordance between the profiles; however, the MLVA method produced an additional four unique profiles amongst the subset of 30 isolates tested. Given that the improved MLVA method was found to be superior to the original MLVA method, it was subsequently used to redefine the molecular epidemiology of L. interrogans serovar Australis in Queensland, Australia. Using cluster analysis, the authors were able to demonstrate clonal links amongst rodent isolates, rodent and human isolates, and rodent and canine isolates. These results highlight the role of rodents in the disease, and also the potential role of MLVA in defining the molecular epidemiology of L. interrogans.

Abbreviations: FAFLP, fluorescent amplified fragment length polymorphism; FAM, 6-carboxyfluorescein; HEX, 6-hexachlorofluorescein; HGDI, Hunter–Gaston diversity index; Ia, index of association; MLVA, multiple-locus variable number of tandem repeats analysis; MLST, multilocus sequence typing; MST, minimum spanning tree; MVT, MLVA type; NED, 2,7,8-benzo-5-fluoro-2,4,7-trichloro-5-carboxyfluorescein; SLV, single-locus variant; VNTR, variable number of tandem repeat.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
INT J SYST EVOL MICROBIOL J MED MICROBIOL MICROBIOLOGY J GEN VIROL ALL SGM JOURNALS
Copyright © 2006 Society for General Microbiology.