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. 1995 Apr;33(4):962–967. doi: 10.1128/jcm.33.4.962-967.1995

Microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species in blood.

S Fujita 1, B A Lasker 1, T J Lott 1, E Reiss 1, C J Morrison 1
PMCID: PMC228076  PMID: 7790469

Abstract

We developed a microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species. Nucleotide sequences derived from the internal transcribed spacer (ITS) region of fungal rDNA were used to develop species-specific oligonucleotide probes for Candida albicans, C. tropicalis, C. parapsilosis, and C. krusei. No cross-hybridization was detected with any other fungal, bacterial, or human DNAs tested. In contrast, a C. (Torulopsis) glabrata probe cross-reacted with Saccharomyces cerevisiae DNA but with no other DNAs tested. Genomic DNA purified from C. albicans blastoconidia suspended in blood was amplified by PCR with fungus-specific universal primers ITS3 and ITS4. With the C. albicans-specific probe labeled with digoxigenin, a biotinylated capture probe, and streptavidin-coated microtitration plates, amplified DNA from a few as two C. albicans cells per 0.2 ml of blood could be detected by enzyme immunoassay.

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Selected References

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