Identification of Genotypically Diverse
            <i>Cryptococcus neoformans</i>
            and
            <i>Cryptococcus gattii</i>
            Isolates by Luminex xMAP Technology

Bovers, Marjan; Diaz, Mara R.; Hagen, Ferry; Spanjaard, Lodewijk; Duim, Birgitta; Visser, Caroline E.; Hoogveld, Hans L.; Scharringa, Jelle; Hoepelman, I. M.; Fell, Jack W.; Boekhout, Teun

doi:10.1128/jcm.00223-07

Cited by 54 publications

(48 citation statements)

References 30 publications

(36 reference statements)

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“…If differentiation within the C. neoformans sp. complex is warranted, Pyrosequencing analysis or cycle sequencing of the rDNA will not adequately resolve these species unless sequencing of a "fast-evolving" region (such as the intergenic spacer region-IGS) is used for this purpose [24,27].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species

Montero

Shea

Jones

et al. 2008

Eur J Clin Microbiol Infect Dis

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Pyrosequencing was used to identify 133 isolates of clinically relevant non-dematiaceous yeasts. These included 97 ATCC strains (42 type strains), 7 UAMH strains, and 29 clinical isolates. Isolates belonged to the following genera: Candida (18 species), Trichosporon (10), Cryptococcus (7), Malassezia (3), Rhodotorula (2), Geotrichum (1), Blastoschizomyces (1) and Kodamaea (1). Amplicons were obtained of a hypervariable ITS region and analyzed using Pyrosequencing technology. The data were evaluated by a BLAST search against the GenBank database and correlated with data obtained by conventional cycle sequencing of the ITS1-5.8S-ITS2 region. Cycle sequencing identified (78.9%) if the isolates to the species level. Pyrosequencing technology identified 69.1%. In 90.1% of all the strains tested, identification results of both sequencing methods were identical. Most Candida isolates can be identified to the species level by pyrosequencing. Trichosporon species and some Cryptococcus cannot be differentiated at the species level. Pyrosequencing can be used for the reliable identification of most commonly isolated nondematiaceous yeasts, with a reduction of cost per identification compared to conventional sequencing.

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Section: Discussionmentioning

confidence: 99%

Evaluation of Pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species

Montero

Shea

Jones

et al. 2008

Eur J Clin Microbiol Infect Dis

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“…In recent years, microsphere-based technology has become an attractive alternative to DNA sequencing. This technology had been applied for species identification in molds and yeasts (16)(17)(18)(19)(20) and for the identification of single nucleotide polymorphisms (SNPs) (21,22). Here, we describe the adaptation of this technology to identify SNPs in the FKS1 HS1 and FKS2 HS1 domains of C. glabrata which confer in vitro resistance to echinocandins.…”

mentioning

confidence: 99%

Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

et al. 2014

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Echinocandins are the recommended treatment for invasive candidiasis due to Candida glabrata. Resistance to echinocandins is known to be caused by nonsynonymous mutations in the hot spot-1 (HS1) regions of the FKS1 and FKS2 genes, which encode a subunit of the ␤-1,3-glucan synthase, the target of echinocandins. Here, we describe the development of a microsphere-based assay using Luminex MagPix technology to identify mutations in the FKS1 HS1 and FKS2 HS1 domains, which confer in vitro echinocandin resistance in C. glabrata isolates. The assay is rapid and can be performed with high throughput. The assay was validated using 102 isolates that had FKS1 HS1 and FKS2 HS1 domains previously characterized by DNA sequencing. The assay was 100% concordant with DNA sequencing results. The assay was then used for high-throughput screening of 1,032 C. glabrata surveillance isolates. Sixteen new isolates with mutations, including a mutation that was new to our collection (del659F), were identified. This assay provides a rapid and cost-effective way to screen C. glabrata isolates for echinocandin resistance.

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“…Previous studies have shown the utility of Luminex- based multiplex applications for the detection of clinically relevant fungi in clinical specimens and for surveillance (2,4,5,8). However, this is the first extensive report of the utility of a Luminex-based assay with 11 probes for the rapid identification of yeast-like organisms, including C. neoformans, from blood cultures.…”

mentioning

confidence: 88%

Detection of Yeasts in Blood Cultures by the Luminex xTAG Fungal Assay

et al. 2012

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A multiplex-PCR Luminex xMAP bead probe fluid array using xTAG analyte-specific reagents (multiplex xTAG fungal ASR assay) was employed for detection of clinically significant Candida species, Cryptococcus neoformans, Histoplasma capsulatum, and Blastomyces dermatitidis from blood cultures. We tested 132 blood cultures negative (n ‫؍‬ 10) or positive (n ‫؍‬ 97) for yeasts and/or bacteria (n ‫؍‬ 25). The assay showed sensitivity and specificity of 100% and 99%, respectively. The xTAG fungal ASR assay is a rapid assay that allows simultaneous identification of multiple yeast species.

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Identification of Genotypically Diverse Cryptococcus neoformans and Cryptococcus gattii Isolates by Luminex xMAP Technology

Cited by 54 publications

References 30 publications

Evaluation of Pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species

Evaluation of Pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species

Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

Detection of Yeasts in Blood Cultures by the Luminex xTAG Fungal Assay

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