Quantitative analysis of nucleic Acid hybridization on magnetic particles and quantum dot-based probes.

Sensors
Sun Hee LimAlexey Dan Chin Yu

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.

References

Jun 21, 1989·Journal of Immunological Methods·F A OssendorpM De Boer
Feb 1, 1984·Human Pathology·J M Hicks
Aug 10, 2000·Analytical Biochemistry·S H SheikhA Mulchandani
Jan 29, 2002·Nucleic Acids Research·A W PetersonR M Georgiadis
Apr 19, 2002·Journal of the American Chemical Society·Joseph WangRonen Polsky
Nov 13, 2004·Angewandte Chemie·Eugenii Katz, Itamar Willner
Aug 11, 2005·Journal of the American Chemical Society·Rumiana BakalovaYoshinobu Baba
Apr 26, 2007·Journal of Agricultural and Food Chemistry·Amit AgrawalShuming Nie

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Citations

Sep 1, 2011·The Analyst·Xin HuangShuifang Zhu
Aug 4, 2010·Analytica Chimica Acta·K Aguilar-ArteagaE Barrado
Mar 25, 2011·Chemistry : a European Journal·Po-Jung Jimmy Huang, Juewen Liu
Sep 30, 2010·Biotechnology Progress·Sara PeetersMarc Van Ranst
Mar 14, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Jiri KudrRene Kizek

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Methods Mentioned

BETA
FRET
flow
fluorescence microscopy
flow cytometry
Assay
electrophoresis
fluorescence

Software Mentioned

ImageJ
BD FACSDiVa
RAS

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