UNIT 13.9 Multiplexed Detection of Fungal Nucleic Acid Signatures
Published Online: 1 APR 2008
Copyright © 2008 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cytometry
How to Cite
Diaz, M. R., Dunbar, S. A. and Jacobson, J. W. 2008. Multiplexed Detection of Fungal Nucleic Acid Signatures. Current Protocols in Cytometry. 44:13.9:13.9.1–13.9.21.
- Published Online: 1 APR 2008
- Published Print: APR 2008
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. Curr. Protocol. Cytom. 44:13.9.1-13.9.21. © 2008 by John Wiley & Sons, Inc.
- bead suspension array;
- nucleic acid;