Development of a nanoparticle-labeled microfluidic immunoassay for detection of pathogenic microorganisms

被引:39
作者
Lin, FYH
Sabri, M
Alirezaie, J
Li, DQ
Sherman, PM
机构
[1] Hosp Sick Children, Inst Res, Div Gastroenterol & Nutr, Toronto, ON M5G 1X8, Canada
[2] Univ Toronto, Dept Mech & Ind Engn, Toronto, ON, Canada
[3] Univ Toronto, Dept Pediat & Lab Med & Pathobiol, Toronto, ON, Canada
[4] Ryerson Univ, Dept Elect & Comp Engn, Toronto, ON, Canada
[5] Univ Waterloo, Dept Syst Design Engn, Waterloo, ON N2L 3G1, Canada
关键词
D O I
10.1128/CDLI.12.3.418-425.2005
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The light-scattering properties of submicroscopic metal particles ranging from 40 to 120 nm in diameter have recently been investigated. These particles scatter incident white light to generate monochromatic light, which can be seen either by the naked eye or by dark-field microscopy. The nanoparticles are well suited for detection in microchannel-based immunoassays. The goal of the present study was to detect Helicobacter pylori-and Escherichia coli O157:H7-specific antigens with biotinylated polyclonal antibodies. Gold particles (diameter, 80 nm) functionalized with a secondary antibiotin antibody were then used as the readout. A dark-field stereomicroscope was used for particle visualization in poly(dimethylsiloxane) microchannels. A colorimetric quantification scheme was developed for the detection of the visual color changes resulting from immune reactions in the microchannels. The microchannel inummoassays reliably detected H. pylori and E. coli O157:117 antigens in quantities on the order of 10 ng, which provides a sensitivity of detection comparable to those of conventional dot blot assays. In addition, the nanoparticles within the microchannels can be stored for at least 8 months without a loss of signal intensity. This strategy provides a means for the detection of nanoparticles in microchannels without the use of sophisticated equipment. In addition, the approach has the potential for use for further miniaturization of immunoassays and can be used for long-term archiving of immunoassays.
引用
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页码:418 / 425
页数:8
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