Branched DNA Signal Amplification for Direct Quantitation of Nucleic Acid Sequences in Clinical Specimens

This chapter provides an overview of basic concepts, principles, and applications of branched DNA (bDNA). The chapter has reviewed the development of bDNA as a method for quantitation of nucleic acid targets and the application of this technology to the study of infectious diseases and cell biology. It begins with the introduction about bDNA. Then goes on to discuss about bDNA in detail giving its overview, sample collection and preparation, target probes, bDNA amplifiers, preamplifiers, labeled probe, probes with non-natural bases, molecular standards, assay requirements, and data analysis. Signal versus target amplification systems is also discussed in the chapter. Infectious disease applications—namely, hepatitis B virus, hepatitis C virus, hepatitis G virus/GB virus C, human immunodeficiency virus type 1, cytomegalovirus, and trypanosoma brucei spp. are discussed next in the chapter. The chapter then turns to discussion of quantitation of messenger RNA. Finally the chapter closes with a summary stating bDNA has several characteristics that distinguish it from the quantitative target amplification systems, including better tolerance of target sequence variability, more direct measurement of target, simpler sample preparation, and less sample-to- sample variation.