Multilocus Sequence Typing of
            <i>Candidaglabrata</i>
            Reveals Geographically EnrichedClades

Dodgson, Andrew; Pujol, Claude; Denning, David W.; Soll, David R.; Fox, Andrew

doi:10.1128/jcm.41.12.5709-5717.2003

Cited by 169 publications

(186 citation statements)

References 46 publications

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“…However, the distribution of alleles was irregular and several alleles were prominent, since three multilocus genotypes represented 52% of the isolates studied. Other studies using microsatellite or MLST data have highlighted the fact that distinct genetic clades of C. glabrata prevail in different geographical regions (8,9,12). Our unrelated isolates were collected from patients from a restricted geographical area, which could partially explain the unequal distribution of genotypes in the population studied.…”

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confidence: 98%

“…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.…”

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confidence: 99%

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Multiple-Locus Variable-Number Tandem-Repeat Analysis for Rapid Typing of Candida glabrata

et al. 2007

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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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confidence: 99%

Multiple-Locus Variable-Number Tandem-Repeat Analysis for Rapid Typing of Candida glabrata

et al. 2007

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“…There are reports about its application for several Candida species (86,116). Recently, two new techniques arouse: amplified fragment length polymorphism (AFLP) (3) and multilocus sequence typing (MLST) (12,32,115).…”

Section: Molecular Biologymentioning

confidence: 99%

Oral Candida spp. colonization in human immunodeficiency virus-infected individuals

Moris¹,

Melhem²,

Martins³

et al. 2008

J. Venom. Anim. Toxins incl. Trop. Dis

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“…Dodgson, et al [58] developed MLST for C. glabrata through amplification and sequencing of fragments from the coding regions of six genes (FKS, LEU2, NMT1, TRP1, UGP1, and URA3). An MLST analysis of 230 C. glabrata isolates from five populations that differed both geographically and temporally confirmed that using the six loci, it was possible to assess genetic diversity and differentiation among isolates of this species [59].…”

Section: Molecular Methods Developed For the Identification And Charamentioning

confidence: 99%

Molecular Methods Developed for the Identification and Characterization of Candida Species

Grisolia¹

2017

IJGS

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Candida species are commensally microorganisms in healthy individuals, but is also the most opportunistic fungal pathogen of human. From benign skin-mucosal forms to invasive ones, Candida infection, can compromise various organs systematic and cause diseases in immunocompromised or critically ill patients. Hence the accurate identification and characterization of the disease-causing strains is crucial for diagnosis, clinical treatment and epidemiological studies of candidacies. Molecular methods the identification and characterization support gives in epidemiological research and in development of novel antifungal with specific molecular targets, especially in the case of resistant Candida species. This review aims to describe the main methods used in the identification and characterization the Candida species and discusses future perspectives.

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Multilocus Sequence Typing of Candidaglabrata Reveals Geographically EnrichedClades

Cited by 169 publications

References 46 publications

Multiple-Locus Variable-Number Tandem-Repeat Analysis for Rapid Typing of Candida glabrata

Multiple-Locus Variable-Number Tandem-Repeat Analysis for Rapid Typing of Candida glabrata

Oral Candida spp. colonization in human immunodeficiency virus-infected individuals

Molecular Methods Developed for the Identification and Characterization of Candida Species

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