Current Microbiology

, Volume 62, Issue 2, pp 518–524 | Cite as

Evaluation of the Use of Selective PCR Amplification of LPS Biosynthesis Genes for Molecular Typing of Leptospira at the Serovar Level

  • Josefa Bezerra da Silva
  • Eneas Carvalho
  • Rudy A. Hartskeerl
  • Paulo L. HoEmail author


Leptospirosis is an important epidemic zoonosis worldwide. Currently, there are more than 250 Leptospira pathogenic serovars known that can potentially infect humans. Conventional classification of leptospires with the serovar as the basic taxon, based on serological recognition of lipopolysaccharide (LPS) composition does not correlate well with species determination, based on general genomic features. Here, we investigate the selective amplification of polymorphic regions from the LPS biosynthesis loci (rfb) as a potential tool for serovar typing of Leptospira interrogans species. Eight pairs of primers were designed to target six ORFs from the rfb operon with varying levels of sequence polymorphism. They were tested both separately and multiplexed. Half of these primer pairs produced serovar-specific amplicons, allowing the identification of some specific serovars and also groups of serovars. It was shown that the serovar classification of Leptospira can be accessed by selective amplification of rfb operons in some cases, which may permit a parallel between the serological and the genomic classifications of Leptospira. As a conclusion, the selective amplification of rfb generated promising and already useful results, but it appears necessary to characterize a larger variety of Leptospira genomes or rfb operons to fully develop this method.


Multiplex Polymerase Chain Reaction Leptospirosis Naam Serological Classification Predict Amplicon Size 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank FAPESP, CNPq, and Fundação Butantan for financial support.

Supplementary material

284_2010_9738_MOESM1_ESM.docx (446 kb)
Individual PCR amplification of LPS biosynthesis loci fragments using a subset of four primer pairs (pair 37–420, 10–268, 14–201, and 37–110), across several L. interrogans serovars. The use of these primer pairs produced amplicons in serovars in which it was not predicted to occur (according to the alignments performed) and also non-specific amplicons, whose size are incompatible with the expected amplicon size (indicated by an arrow). Serovars in which unpredicted amplicons occurred are presented in bold, and serovars in which non-specific amplicons occurred are italicized; otherwise, when the predicted result was obtained for the serovar, it was underlined. An approximate size marker (bp) is shown for each figure subset. Serovar Hardjo belongs to subtype Hardjoprajitno. (DOCX 446 kb)


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Josefa Bezerra da Silva
    • 1
  • Eneas Carvalho
    • 1
  • Rudy A. Hartskeerl
    • 2
  • Paulo L. Ho
    • 1
    Email author
  1. 1.Centro de BiotecnologiaInstituto ButantanSão PauloBrazil
  2. 2.KIT Biomedical ResearchRoyal Tropical InstituteAmsterdamThe Netherlands

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