Unique oligonucleotide primers in PCR for identification of
Cryptococcus neoformans.
T. G. Mitchell, E. Z. Freedman, T. J. White & J. W. Taylor
Department of Microbiology, Duke University Medical Center,
Durham, North Carolina 27710.
J Clin Microbiol 32: 253-255 (1994) [94172038]
Abstract
On the basis of a comparison of rRNA sequences coding the internal
transcribed spacer (ITS) and 5.8S regions of Filobasidiella
neoformans, the teleomorph of Cryptococcus neoformans,
and members of its most closely related taxa, unique
oligonucleotides were designed for the specific amplification of
DNA only from C. neoformans. Various combinations of these
primers, designated CN4, CN5, and CN6, and the previously
described fungal primers ITS1 and ITS2 were tested in PCR for
their ability to amplify DNA from 37 strains of C.
neoformans and 31 other isolates representing 18 species of
yeasts. The combination of primers CN4 and CN5 amplified DNA from
both varieties of C. neoformans but from none of the other
species tested. Other pairs of primers (namely, CN5-CN6, CN4-ITS1,
and CN6-ITS1) amplified DNA only from C. neoformans and
from the saprophyte Filobasidiella depauperata, which is
the only other member of the genus Filobasidiella. With
appropriate controls, these specific oligonucleotides may be used
as primers in PCR to identify C. neoformans.