Nucleic Acid Amplification and Detection Methods

Abstract

Nucleic acid detection methods play an increasingly important role in the detection of viral infection. This chapter describes the major nucleic acid testing methods and assists in test selection. Nucleic acid amplification methods are classified as target or probe amplification methods based upon the source of the nucleic acid that is amplified in the procedure. Target amplification methods are among the oldest and best characterized nucleic acid amplification methodologies. Nucleic acid amplification detection methods include polymerase chain reaction (PCR), nucleic acid sequence-based amplification (NASBA) and transcription-mediated amplification (TMA). Nucleic acid detection methods include SYBR green, fluorescence resonance energy transfer (FRET) system and hydrolysis (TaqMan) probes. Other amplification methods described in the chapter are part of closed assay systems whose manufacturers discourage or prohibit in-house development. Nucleic acid detection methods have also made significant improvements in one's ability to detect fastidious and slow-growing viruses (e.g., human parvovirus, Epstein-Barr virus, and certain enteroviruses), viruses that are dangerous to amplify in culture (e.g., human immunodeficiency virus and certain hemorrhagic fever viruses), and viruses that are present in low concentrations. While nucleic acid detection methods will never completely replace culture and direct fluorescent antibody methods, nucleic acid detection methods will continue to play an important role in the detection and monitoring of viral diseases.