Nucleic Acid Amplification Technologies: Application to Disease Diagnosis

By Stephen A. Morse and Helen Lee

The polymerase chain reaction (PCR) has proved to be a powerful and versatile tool and has opened new avenues in molecular biol- ogy. Alternative nucleic acid amplification techniques, such as the ligase chain reaction (LCR), nucleic acid sequence-based amplifica- tion (NASBA), and transcription-mediated amplification (TMA), a variation of NASBA, are also now available. These techniques are all designed to amplify specific nucleic acid sequences in an expo- nential manner, thus providing a basis for extremely sensitive diag- nostic assays. However, despite the widespread and successful ap- plication of genomic amplification techniques in biological research, they have not yet reached the point of routine use in clini- cal laboratories. Thus, although the R&D investment in nucleic acid diagnostics is in excess of $250 million annually, clinical ap- plications remain relatively modest. One of the principal reasons for this delay in clinical application has been the problem of acci- dental contamination of negative clinical specimens with minute amounts of amplified products from a previous positive reaction. Carry-over contamination of amplicons can now be prevented by chemical means or the use of a closed reaction system. However, the current instrumentation is essentially modular in nature, com- prising machines that perform the three essential steps of nucleic acid amplification technology: sample preparation, the amplifica- tion reaction, and detection of products. Consequently, the test pro- cedures are more complicated with somewhat lower sample throughput than the enzyme immunoassays currently performed in clinical laboratories.

• Format: Hardcover
• Language: English
• ISBN: 0817639217
• ISBN13: 9780817639211
• Release Date: July 1997
• Publisher: Birkhauser
• Length: 286 Pages