Characteristics and Applications of Nucleic Acid Sequence-Based Amplification (NASBA)
- 1 January 2002
- journal article
- research article
- Published by Springer Science and Business Media LLC in Molecular Biotechnology
- Vol. 20 (2), 163-180
- https://doi.org/10.1385/mb:20:2:163
Abstract
Nucleic acid sequence-based amplification (NASBA) is a sensitive, isothermal, transcription-based amplification system specifically designed for the detection of RNA targets. In some NASBA systems, DNA is also amplified though very inefficiently and only in the absence of the corresponding RNA target or in case of an excess (>1,000-fold) of target DNA over RNA. As NASBA is primer-dependent and amplicon detection is based on probe binding, primer and probe design rules are included. An overview of various target nucleic acids that have been amplified successfully using NASBA is presented. For the isolation of nucleic acids prior to NASBA, the "Boom" method, based on the denaturing properties of guanidine isothiocyanate and binding of nucleic acid to silica particles, is preferred. Currently, electro-chemiluminescence (ECL) is recommended for the detection of the amplicon at the end of amplification. In the near future, molecular beacons will be introduced enabling "real-time detection," i.e., amplicon detection during amplification. Quantitative HIV-1 NASBA and detection of up to 48 samples can then be performed in only 90 min.Keywords
This publication has 58 references indexed in Scilit:
- Expression Dynamics of Human Cytomegalovirus Immune Evasion GenesUS3, US6,andUS11in the Blood of Lung Transplant RecipientsThe Journal of Infectious Diseases, 2001
- Nucleic Acid Sequence-Based Amplification of Aspergillus RNA in Blood SamplesJournal of Clinical Microbiology, 2001
- Evaluation of the NucliSens Basic Kit for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Genital Tract Specimens Using Nucleic Acid Sequence-Based Amplification of 16S rRNAJournal of Clinical Microbiology, 2001
- MACROPHAGE-DERIVED CHEMOKINE GENE EXPRESSION IN HUMAN AND MACAQUE CELLS: mRNA QUANTIFICATION USING NASBA TECHNOLOGYCytokine, 2001
- A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytesNucleic Acids Research, 1999
- Typing ofMycoplasma pneumoniaeby nucleic acid sequence-based amplification, NASBA®Molecular and Cellular Probes, 1996
- Serum HCV RNA levels assessed by quantitative NASBA®: stability of viral load over time, and lack of correlation with liver diseaseJournal of Hepatology, 1996
- Molecular Beacons: Probes that Fluoresce upon HybridizationNature Biotechnology, 1996
- Nucleic acid sequence-based amplification (NASBA) for the identification of mycobacteriaJournal of General Microbiology, 1993
- Self-sustained sequence replication (3SR): an isothermal transcription-based amplification system alternative to PCR.Genome Research, 1991