Single-Molecule Immunoassay and DNA Diagnoasis
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.
Y. Ma et al., "Single-Molecule Immunoassay and DNA Diagnoasis," Electrophoresis, Wiley-VCH, Jan 2001.
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
Article - Journal
© 2001 Wiley-VCH, All rights reserved.