[Detection of nucleic acid sequences by bioluminescence. Importance of an internal reference system for qualitative and quantitative analysis].
Various luminescent detection systems were used to detect and quantify simultaneously several probes. For quantitative analysis, one probe was used as an internal standard to evaluate the amplification yield and the others to quantify the amplified target. After using hybridization in solution, accurate results were obtained. For a qualitative analysis, amplified sequences were hybridized to specific oligonucleotides which were labelled or immobilized. To compare amplified fragments to products of known sequences, we developed a more efficient technique based on DNA strand exchange occurring during hybridization in solution. This method allowed to detect a single base substitution in DNA fragments of 280 base pairs.
Nicolas
J C
JC
Inserm U 58, 60, Montpellier, France.
Térouanne
B
B
Balaguer
P
P
Chikhaoui
Y
Y
Defacque
H
H
Boussioux
A M
AM
D001483:Base Sequence; D004279:DNA, Viral / Q000737:chemistry; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012015:Reference Standards
D004279:DNA, Viral
Détection par bioluminescence de séquences d'acides nucléiques. Importance du système de référence interne pour l'analyse qualitative et quantitative.