Heterologous system for the detection of chemically-labeled...

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167/37, 150/8.5

G01N 33/535 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01)

Patent

CA 1314810

HETEROLOGOUS SYSTEM FOR THE DETECTION OF CHEMICALLY- LABELED DNA AND OTHER BIOLOGICAL MATERIALS PROVIDING A RECEPTOR OR TARGET MOIETY THEREON ABSTRACT OF THE DISCLOSURE The detection and/or identification of materials of interest, such as biological materials, e.g. chemically labeled DNA, which provide a receptor thereon, is improved by employing a heterologous detection system by employing more than one binding reactions, such as reactions involving the affinity of lectin to a glyco- protein or sugar group and the affinity between biotin and avidin or streptavidin, avidin itself being a glycoprotein. Particularly useful in the heterologous detection system are biotinylated lectins, such as biotinylated Concanavalin A, biotinylated dextran, lectin- glycoprotein complexes, such as lectin-avidin and lectin- enzyme complexes and streptavidin-biotinylated enzyme complexes. By way of example, the detection of single- stranded glycosylated DNA hybridized to complementary single-stranded DNA is detected by bringing the hybridized doudle-stranded DNA into contact with a lectin, such as Concanavalin A, for attachment to the glycosyl group of the glycosylated DNA, followed by contact with a biotinylated dextran, a polysaccharide, and then by contact with a lectin-enzyme complex, such as Concanavalin A-horseradish peroxidase or acid phosphatase. Also, there would be employed an enzyme complex compris- ing streptavidin-biotinylated horseradish peroxidase. The lectin-enzyme complex would attach itself to the sugar or saccharide of the biotinylated dextran which is also attached to the lectin which is fixed to the glycosyl groups of the hybridized DNA. The streptavidin- biotinylated enzyme complex would attach itself to the biotin moieties of the biotinylated dextran which, as indicated, is linked to the lectin attached to the glycosyl groups of the target double-stranded DNA.

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