Affiliation:
1. Mycotic Diseases Branch, Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia
Abstract
ABSTRACT
Specific oligonucleotide probes were developed to identify medically important fungi that display yeast-like morphology in vivo. Universal fungal primers ITS1 and ITS4, directed to the conserved regions of ribosomal DNA, were used to amplify DNA from
Histoplasma capsulatum
,
Blastomyces dermatitidis
,
Coccidioides immitis
,
Paracoccidioides brasiliensis
,
Penicillium marneffei
,
Sporothrix schenckii
,
Cryptococcus neoformans
, five
Candida
species, and
Pneumocystis carinii
. Specific oligonucleotide probes to identify these fungi, as well as a probe to detect all dimorphic, systemic pathogens, were developed. PCR amplicons were detected colorimetrically in an enzyme immunoassay format. The dimorphic probe hybridized with DNA from
H
.
capsulatum
,
B
.
dermatitidis
,
C
.
immitis
,
P
.
brasiliensis
, and
P
.
marneffei
but not with DNA from nondimorphic fungi. Specific probes for
H
.
capsulatum
,
B
.
dermatitidis
,
C
.
immitis
,
P
.
brasiliensis
,
P
.
marneffei
,
S
.
schenckii
,
C
.
neoformans
, and
P
.
carinii
hybridized with homologous but not heterologous DNA. Minor cross-reactivity was observed for the
B
.
dermititidis
probe used against
C
.
immitis
DNA and for the
H
.
capsulatum
probe used against
Candida albicans
DNA. However, the
C
.
immitis
probe did not cross-react with
B
.
dermititidis
DNA, nor did the dimorphic probe hybridize with
C
.
albicans
DNA. Therefore, these fungi could be differentiated by a process of elimination. In conclusion, probes developed to yeast-like pathogens were found to be highly specific and should prove to be useful in differentiating these organisms in the clinical setting.
Publisher
American Society for Microbiology
Cited by
120 articles.
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