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Journal of Clinical Microbiology, October 2001, p. 3505-3511, Vol. 39, No. 10
Mycotic Diseases Branch, Division of
Bacterial and Mycotic Diseases, Centers for Disease Control and
Prevention, Atlanta, Georgia
Received 31 May 2001/Returned for modification 22 July
2001/Accepted 7 August 2001
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.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.10.3505-3511.2001
Rapid Identification of Dimorphic and Yeast-Like
Fungal Pathogens Using Specific DNA Probes
*
Corresponding author. Mailing address: Mailstop G-11,
CDC, 1600 Clifton Rd., NE, Atlanta, GA 30333. Phone: (404) 639-4340. Fax: (404) 639-3546. E-mail: mil6{at}cdc.gov.
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