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1

Diagnostics, Institut Pasteur Korea, 696 Sampyeong-dong, Bundang-gu, Seongnam City, Gyeonggi-do, 443-270, Korea

2

Cell Biophysics, Institut Pasteur Korea, 696 Sampyeong-dong, Bundang-gu, Seongnam City, Gyeonggi-do, 443-270, Korea

3

Virology Unit, Institut Pasteur in Cambodia, 5, Monivong boulevard, Phnom Penh, Cambodia





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Abstract 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. View Full-Text

Keywords: magnetic particles; quantum dots; hybridization; DNA; flow cytometry; fluorescence microscopy magnetic particles; quantum dots; hybridization; DNA; flow cytometry; fluorescence microscopy





Autor: Sun Hee Lim 1,* , Felix Bestvater 2, Philippe Buchy 3, Sek Mardy 3 and Alexey Dan Chin Yu 1,*

Fuente: http://mdpi.com/



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