Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology

M Bovers; M R Diaz; F Hagen; L Spanjaard; B Duim; C E Visser; H L Hoogveld; J Scharringa; I M Hoepelman; J W Fell; T Boekhout

doi:10.1128/JCM.00223-07

Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology

M Bovers, M R Diaz, F Hagen, L Spanjaard, B Duim, C E Visser, H L Hoogveld, J Scharringa, I M Hoepelman, J W Fell, T Boekhout

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans and Cryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.

Original language	English
Pages (from-to)	1874-83
Number of pages	10
Journal	Journal of Clinical Microbiology
Volume	45
Issue number	6
DOIs	https://doi.org/10.1128/JCM.00223-07
Publication status	Published - Jun 2007

Keywords

Adult
Aged
Cerebrospinal Fluid
Cryptococcosis
Cryptococcus
Cryptococcus neoformans
DNA, Fungal
Female
Flow Cytometry
Genotype
Humans
Male
Meningitis, Cryptococcal
Microspheres
Middle Aged
Molecular Sequence Data
Mycological Typing Techniques
Polymerase Chain Reaction
Sequence Analysis, DNA
Species Specificity
Suspensions

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10.1128/JCM.00223-07

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Bovers, M., Diaz, M. R., Hagen, F., Spanjaard, L., Duim, B., Visser, C. E., Hoogveld, H. L., Scharringa, J., Hoepelman, I. M., Fell, J. W., & Boekhout, T. (2007). Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology. Journal of Clinical Microbiology, 45(6), 1874-83. https://doi.org/10.1128/JCM.00223-07

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title = "Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology",

abstract = "A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans and Cryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.",

keywords = "Adult, Aged, Cerebrospinal Fluid, Cryptococcosis, Cryptococcus, Cryptococcus neoformans, DNA, Fungal, Female, Flow Cytometry, Genotype, Humans, Male, Meningitis, Cryptococcal, Microspheres, Middle Aged, Molecular Sequence Data, Mycological Typing Techniques, Polymerase Chain Reaction, Sequence Analysis, DNA, Species Specificity, Suspensions",

author = "M Bovers and Diaz, {M R} and F Hagen and L Spanjaard and B Duim and Visser, {C E} and Hoogveld, {H L} and J Scharringa and Hoepelman, {I M} and Fell, {J W} and T Boekhout",

year = "2007",

month = jun,

doi = "10.1128/JCM.00223-07",

language = "English",

volume = "45",

pages = "1874--83",

journal = "Journal of Clinical Microbiology",

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publisher = "American Society for Microbiology",

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AU - Bovers, M

AU - Diaz, M R

AU - Hagen, F

AU - Spanjaard, L

AU - Duim, B

AU - Visser, C E

AU - Hoogveld, H L

AU - Scharringa, J

AU - Hoepelman, I M

AU - Fell, J W

AU - Boekhout, T

PY - 2007/6

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N2 - A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans and Cryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.

AB - A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans and Cryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.

KW - Adult

KW - Aged

KW - Cerebrospinal Fluid

KW - Cryptococcosis

KW - Cryptococcus

KW - Cryptococcus neoformans

KW - DNA, Fungal

KW - Female

KW - Flow Cytometry

KW - Genotype

KW - Humans

KW - Male

KW - Meningitis, Cryptococcal

KW - Microspheres

KW - Middle Aged

KW - Molecular Sequence Data

KW - Mycological Typing Techniques

KW - Polymerase Chain Reaction

KW - Sequence Analysis, DNA

KW - Species Specificity

KW - Suspensions

U2 - 10.1128/JCM.00223-07

DO - 10.1128/JCM.00223-07

M3 - Article

C2 - 17442792

SN - 0095-1137

VL - 45

SP - 1874

EP - 1883

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

IS - 6

ER -

Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology

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Keywords

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