Rapid differentiation of Aspergillus species from other medically important opportunistic molds and yeasts by PCR-enzyme immunoassay.
We developed a PCR-based assay to differentiate medically important species of Aspergillus from one another and from other opportunistic molds and yeasts by employing universal, fungus-specific primers and DNA probes in an enzyme immunoassay format (PCR-EIA). Oligonucleotide probes, directed to the internal transcribed spacer 2 region of ribosomal DNA from Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus, Aspergillus ustus, and Aspergillus versicolor, differentiated 41 isolates (3 to 9 each of the respective species; P < 0.001) in a PCR-EIA detection matrix and gave no false-positive reactions with 33 species of Acremonium, Exophiala, Candida, Fusarium, Mucor, Paecilomyces, Penicillium, Rhizopus, Scedosporium, Sporothrix, or other aspergilli tested. A single DNA probe to detect all seven of the most medically important Aspergillus species (A. flavus, A. fumigatus, A. nidulans, A. niger, A. terreus, A. ustus, and A. versicolor) was also designed. Identification of Aspergillus species was accomplished within a single day by the PCR-EIA, and as little as 0.5 pg of fungal DNA could be detected by this system. In addition, fungal DNA extracted from tissues of experimentally infected rabbits was successfully amplified and identified using the PCR-EIA system. This method is simple, rapid, and sensitive for the identification of medically important Aspergillus species and for their differentiation from other opportunistic fungi.
de Aguirre
Liliana
L
Mycotic Diseases Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., NE, Mailstop G-11, Atlanta, GA 30333, USA.
Hurst
Steven F
SF
Choi
Jong Soo
JS
Shin
Jong Hee
JH
Hinrikson
Hans Peter
HP
Morrison
Christine J
CJ
D000818:Animals; D001228:Aspergillosis / Q000175:diagnosis / Q000662:veterinary; D001230:Aspergillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D017931:DNA Primers; D015342:DNA Probes; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D005516:Food Microbiology; D006801:Humans; D007124:Immunoenzyme Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D011817:Rabbits / Q000382:microbiology; D015003:Yeasts / Q000145:classification / Q000235:genetics / Q000302:isolation & purification
D017931:DNA Primers; D015342:DNA Probes; D004271:DNA, Fungal; D004275:DNA, Ribosomal