Currently the universally accepted standard procedure for characterizing and identifying strains of Leishmania is isoenzyme analysis. However, in the Mediterranean area, despite their very wide geographical distribution, most Leishmania infantum strains belong to zymodeme MON-1. In order to increase our understanding of polymorphism in strains of L. infantum, we developed PCR assays amplifying 10 microsatellites and sequenced PCR products.

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PCPs play an important role in travel medicine. This study showed that their high level of knowledge in travel medicine was mostly linked to their motivation to practice in this specialized discipline.

A multiple-locus variable-number tandem-repeat analysis (MLVA) using six microsatellite markers was assessed in 127 Candida glabrata isolates. Thirty-seven different genotypes, stable both in vitro and in vivo, were observed. The highest discriminatory power (D ‫؍‬ 0.902) was reached by using only four markers. MLVA seems to be relevant for C. glabrata typing.Candida glabrata has recently emerged as a major pathogen, causing mucosal and systemic infections (10,17). Several methods, such as electrophoretic karyotyping, restriction enzyme analysis, Southern blotting with probes, randomly amplified polymorphic DNA, and multilocus sequence typing (MLST), have been used to distinguish and type C. glabrata isolates (2,7,8,14,21,28). Microsatellite polymorphism analysis has been widely used for typing fungi (3,4,11,15,18,25), and this could be an alternative, easy-to-perform, reproducible method suitable for large-scale studies of C. glabrata epidemiology. Recently, Foulet et al. described three polymorphic microsatellite markers to investigate the delineation of clinical C. glabrata isolates (12). The discriminatory power of this method was good but not optimal. The aim of our study was to assess a microsatellite-based multiple-locus variable-number tandemrepeat analysis (MLVA) using new markers for C. glabrata typing.One hundred twenty-seven C. glabrata strains were analyzed, including four reference strains, 98 independent clinical isolates, and 25 epidemiologically related isolates. These 25 were from the blood cultures and peripheral site isolates of eight patients with C. glabrata candidemia. Genomic DNA was extracted by boiling with Chelex resin as previously described (6, 23). Six microsatellite markers were selected from the C. glabrata DNA sequences available in the GenBank database (29). Primer sequences were designed with Primer3 software (24), and locations in the C. glabrata genome were determined with the Genolevures database (http://cbi.labri.fr/Genolevures/) ( Table 1). For each primer set, PCRs were performed with a 20-l final volume containing 1 l of DNA, each deoxynucleoside triphosphate at 200 M, a forward primer and a 5Ј-dyelabeled reverse primer at 0.25 M each, and 1 U of Taq DNA polymerase (Promega, Madison, WI). The amplification conditions were 5 min at 95°C, followed by 35 cycles of 95°C for 30 s, 52°C for 30 s, and 72°C for 45 s and then a final step of 72°C for 10 min. For any given isolate, amplicons of each PCR were pooled before multiplex fragment sizing with a Ceq 8000 Genetic Analyzer (Beckman Coulter, Fullerton, CA). Strain IHEM 9670 was run as a control in each experiment. As allele sizing depends on dyes and the analyzer used for electrophoresis, results were expressed as the exact size of the sequence (determined by sequencing of each representative allele for each locus) to allow further interlaboratory comparisons of MLVA results (13,20). The reproducibility and stability of the method were assessed as described elsewhere (25) . Primer specificity was also tested against the sequ...

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