Potential application of low-stringency single specific primer-PCR in the identification of Leptospira in the serum of patients with suspected leptospirosis

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Abstract:

In this study we tested the potential use of low-stringency single specific primer-PCR (LSSP-PCR) for genetically typing Leptospira directly from biological samples. Serum samples obtained from 29 patients with clinically suspected leptospirosis were amplified by specific PCR, using the previously selected G1 and G2 primers. The PCR products of approximately 300 bp were subsequently used as a template for LSSP-PCR analysis. We were able to produce genetic signatures from the leptospires present in the human samples, which permitted us to make a preliminary identification of the infective serovar by comparing the LSSP-PCR profiles obtained directly from serum samples with those from reference leptospires. Thus, LSSP-PCR has the potential to become a useful diagnostic tool for identifying leptospires in biological samples without the need for bacteria isolation and culture.Key words: LSSP-PCR, Leptospira.

Dans cette étude, nous avons analysé l'utilisation potentielle du PCR-LSSP (low-stringency single specific primer, amorces simples spécifiques à basse sélectivité) pour le typage génétique de Leptospira directement à partir d'échantillons biologiques. Des échantillons de sérum obtenus de 29 patients soupçonnés d'être atteints de leptospirose clinique ont été amplifiés par PCR spécifique en employant les amorces G1 et G2 préalablement sélectionnées. Les produits de PCR d'environ 300 pb ont été par la suite utilisés pour l'amplification par PCR-LSSP. Nous avons été capables de générer des signatures génétiques des leptospires présents dans les échantillons humains, qui nous ont permis d'effectuer une identification préliminaire des sérovars infectieux en comparant leurs profils de PCR-LSSP obtenus directement des échantillons de sérums avec ceux de leptospires de référence. Ainsi, le PCR-LSSP pourrait devenir un outil de diagnostic utile pour l'identification de leptospires dans des échantillons biologiques, sans avoir recours à l'isolation et à la culture de bactéries.Mots clés : PCR-LSSP, Leptospira.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: December 1, 2004

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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