In the present study we describe sandwich design hybridization probes consisting of magnetic particles
(MP) and quantum dots (QD) with target DNA, and their application in the detection of avian influenza
virus (H5N1) sequences. Hybridization of 25-, 40-, and 100-mer target DNA with both probes was analyzed
and quantified by flow cytometry and fluorescence microscopy on the scale of single particles. The
following steps were used in the assay: (i) target selection by MP probes and (ii) target detection by QD
probes. Hybridization efficiency between MP conjugated probes and target DNA hybrids was controlled by
a fluorescent dye specific for nucleic acids. Fluorescence was detected by flow cytometry to distinguish
differences in oligo sequences as short as 25-mer capturing in target DNA and by gel-electrophoresis in the
case of QD probes. This report shows that effective manipulation and control of micro- and nanoparticles in
hybridization assays is possible.
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