Quantitative analysis of specific mRNA transcripts using a competitive PCR assay with electrochemiluminescent detection.

Reverse transcriptase-polymerase chain reaction (RT--PCR) has been widely utilized for both the qualitative and quantitative assessment of the levels of specific mRNA transcripts in living systems. Quantitation of specific transcripts has often proved to be problematic because of the difficulty associated with relating the PCR-amplified product to the starting cDNA representing the mRNA of interest. We have overcome these difficulties and have developed a competitive PCR assay employing the property of electrochemiluminescence for the detection of PCR products. This assay possesses the dual advantage of being both nonradioactive and highly sensitive.