Branched DNA (bDNA) Technology for Direct Quantification of Nucleic Acids: Design and Performance

Abstract

The branched DNA (bDNA) assay represents a significant advance in the direct quantification of nucleic acid molecules for research and clinical applications. Other methods for the detection and quantification of nucleic acid molecules, such as polymerase chain reaction (PCR) and nucleic acid sequence-based amplification (NASBA), involve molecular amplification of target nucleic acids followed by detection of amplified products. While these target amplification methods can be extremely sensitive, they suffer from two major disadvantages. First, it is difficult to obtain reproducible quantification since the target nucleic acid must be extracted and amplified in order to measure it. Second, numerous precautions must be taken to avoid false positive results caused by contamination of samples with PCR products and carryover from other specimens. A new departure from target amplification methods, the bDNA assay, directly measures nucleic acid molecules by boosting the reporter signal, rather than by replicating target sequences as the means of detection (Figure 1), and hence it is not subject to the errors inherent in the extraction and amplification of target nucleic acids. An ideal tool for nucleic acid quantification, the bDNA assay detects nucleic acid molecules at their physiological concentration, yields highly reproducible quantification, and eliminates false positives due to contamination.KeywordsTarget ProbeDirect QuantificationTarget Nucleic AcidNucleic Acid MoleculeReproducible QuantificationThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.