title: Detection of Candida albicans DNA from blood samples using a novel electrochemical assay.
abstract: The genus Candida contains a number of yeast species which are opportunistic pathogens and are associated with life-threatening infections in immunocompromised individuals. Provision of appropriate therapy relies on the rapid identification of the infecting species, and existing methods of identifying Candida species in clinical samples are time and resource intensive and are not always specific enough to differentiate between drug-susceptible and drug-resistant species. We have previously developed a system for the rapid detection of yeast pathogens in clinical samples using PCR followed by hybridization with a suite of five species-specific, electrochemically labelled DNA probes. The limit of detection of the assay was shown to be 37 fg (∼1 genome) per reaction using extracted genomic DNA. We carried out a study to test the limit of detection of one of the probes, CA PR3, using blood samples from a healthy donor that were spiked with genomic DNA or with C. albicans cells. Our results demonstrated a limit of detection of 37 fg (ml blood)(-1) (∼1 genome ml(-1)) using extracted DNA or 10 c.f.u. (ml blood)(-1) using C. albicans cells, indicating that the assay is capable of detecting C. albicans nucleic acid at levels that are encountered in clinical samples.
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, UK.
authors.forename: Alastair
authors.initials: A
authors.lastname: Muir
authors.affiliation: Atlas Genetics Ltd, White Horse Industrial Park, Trowbridge, UK.
authors.forename: Gordon
authors.initials: G
authors.lastname: Forrest
authors.affiliation: Atlas Genetics Ltd, White Horse Industrial Park, Trowbridge, UK.
authors.forename: John
authors.initials: J
authors.lastname: Clarkson
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Wheals
mesh_terms: D001769:Blood / Q000382:microbiology*; D002176:Candida albicans / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004271:DNA, Fungal / Q000097:blood*; D055664:Electrochemical Techniques / Q000379:methods*; D006801:Humans; D009172:Mycology / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D004271:DNA, Fungal