Real-time nucleic acid sequence-based amplification in nanoliter volumes.
Real-time nucleic acid sequence-based amplification (NASBA) is an isothermal method specifically designed for amplification of RNA. Fluorescent molecular beacon probes enable real-time monitoring of the amplification process. Successful identification, utilizing the real-time NASBA technology, was performed on a microchip with oligonucleotides at a concentration of 1.0 and 0.1 microM, in 10- and 50-nL reaction chambers, respectively. The microchip was developed in a silicon-glass structure. An instrument providing thermal control and an optical detection system was built for amplification readout. Experimental results demonstrate distinct amplification processes. Miniaturized real-time NASBA in microchips makes high-throughput diagnostics of bacteria, viruses, and cancer markers possible, at reduced cost and without contamination.
Gulliksen
Anja
A
NorChip AS, Industriveien 8, 3490 Klokkarstua, Norway. Anja.Gulliksen@norchip.com
Solli
Lars
L
Karlsen
Frank
F
Rogne
Henrik
H
Hovig
Eivind
E
Nordstrøm
Trine
T
Sirevåg
Reidun
R
D006801:Humans; D036103:Nanotechnology / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D021161:Self-Sustained Sequence Replication / Q000379:methods*