Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology.
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.
Bovers
M
M
CBS-Fungal Biodiversity Centre, Uppsalalaan 8, NL-3584CT Utrecht, The Netherlands.
Diaz
M R
MR
Hagen
F
F
Spanjaard
L
L
Duim
B
B
Visser
C E
CE
Hoogveld
H L
HL
Scharringa
J
J
Hoepelman
I M
IM
Fell
J W
JW
Boekhout
T
T
D000328:Adult; D000368:Aged; D002555:Cerebrospinal Fluid / Q000382:microbiology; D003453:Cryptococcosis / Q000382:microbiology; D003454:Cryptococcus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D003455:Cryptococcus neoformans / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005838:Genotype; D006801:Humans; D008297:Male; D016919:Meningitis, Cryptococcal / Q000382:microbiology; D008863:Microspheres; D008875:Middle Aged; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013535:Suspensions
D004271:DNA, Fungal; D013535:Suspensions