Title

Single-Molecule Immunoassay and DNA Diagnoasis

Abstract

Many assays relevant to disease diagnosis are based on electrophoresis, where the migration velocity is used for distinguishing molecules of different size or charge. However, standard gel electrophoresis is not only slow but also insensitive. We describe a single-molecule imaging procedure to measure the electrophoretic mobilities of up to 100 000 distinct molecules every second. the results correlate well with capillary electrophoresis (CE) experiments and afford confident discrimination between normal (16.5 kbp) and abnormal (6.1 kbp) mitochondrial DNA fragments, or -phycoerythrin-labeled digoxigenin (BP-D) and its immunocomplex (anti-D-BP-D). This demonstrates that virtually all electrophoresis diagnostic protocols from slab gels to CE should be adaptable to single-molecule detection. This opens up the prossibility of screening single copies of DNA or proteins within single biological cells for disease markers without performing polymerase chain reaction (PCR) or other biological amplification.

Department(s)

Chemistry

Sponsor(s)

National Institutes of Health (U.S.)
United States. Department of Energy. Office of Basic Energy Sciences. Division of Chemical Sciences (E.S.Y.)

Keywords and Phrases

Capillary Electrophoresis; DNA; Immunoassay; Single Molecules

Document Type

Article - Journal

Document Version

Final Version

File Type

text

Language(s)

English

Rights

© 2001 Wiley-VCH, All rights reserved.

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