Quantitative polymerase chain reaction and solid-phase capture nucleic acid detection.

ABSTRACT

The combination of PCR amplification and chemiluminescent detection of PCR products provides a highly sensitive system for the quantitation of DNA and RNA. The broad dynamic range of the chemiluminescent detection assay simplifies the selection of cycling and concentration parameters critical to harnessing the quantitative aspects of PCR amplification. Detection of 200 amol of PCR product is attained using the described procedures. The tube or microplate format of the assay avoids many of the limitations associated with other methods of PCR quantitation involving gel electrophoresis. This detection methodology can be applied to a variety of quantitative nucleic acid assays, including viral load and gene expression analysis.