Multiplexed detection of fungal nucleic acid signatures.
Diagnoses of opportunistic mycotic infections constitute an increasing clinical problem. Conventional diagnostic tests are time consuming and lack specificity and sensitivity for accurate and timely prognoses. This unit provides a comprehensive description of a fungal detection method that combines nucleic acid signatures with flow cytometry. The multiplexed assay, which uses xMAP technology, consists of unique fluorescent microspheres covalently bound to species-specific fungal oligonucleotide probes. In the presence of the complementary target sequence, the probe hybridizes to its biotinylated target. Quantification of the reaction is based on the fluorescence signal of the reporter molecule that binds to the biotin moieties of the target. The assay can be expanded to include other microorganisms and has the capability to simultaneously test 100 different fungal probes per tube/well. The speed, flexibility in design, and high-throughput capability makes this assay an attractive diagnostic tool for fungal infections and other related maladies.
Diaz
Mara R
MR
University of Miami, Rosenstiel School of Marine and Atmospheric Science, Miami, Florida, USA.
Dunbar
Sherry A
SA
Jacobson
James W
JW
D001681:Biological Assay; D001710:Biotin / Q000378:metabolism; D019921:Biotinylation; D015342:DNA Probes / Q000235:genetics; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D005434:Flow Cytometry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism; D005658:Fungi / Q000235:genetics*; D006801:Humans; D008863:Microspheres; D009181:Mycoses / Q000175:diagnosis; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000235:genetics; D009894:Opportunistic Infections / Q000175:diagnosis; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors
D015342:DNA Probes; D004271:DNA, Fungal; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D015345:Oligonucleotide Probes; D001710:Biotin