Use of bioluminescence in nucleic acid hybridization reactions

✍ Scribed by Balaguer, Patrick ;Térouanne, Béatrice ;Boussioux, Anne-Marie ;Nicolas, Jean-Claude

• Book ID: 102762027
• Publisher: John Wiley and Sons
• Year: 1989
• Weight: 546 KB
• Volume: 4
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170040142

✦ Synopsis

Luminescence reactions can be used t o detect specific nucleic acid sequences hybridized with a nucleic probe. Different labels such as cytidine sulphone, fluorescein, and biotin can be incorporated into DNA or oligonucleotide molecules and detected by antibody or avidin conjugates coupled to glucose-6P dehydrogenase.

On supports such as nitrocellulose filters, sensitivity is not greatly increased using luminescence, but detection is rapid and easy t o perform using Polaroid film. Moreover, hybridization can be performed with different labelled probes on the same sample.

In solution, luminescence can be used t o monitor sandwich reactions. The method is less sensitive than detection on filters but can easily be automated. The performance of these assays can be increased considerably by enzymatic amplification of the target catalysed by a thermostable polymerase.