C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
195/1.12, 195/35
C12Q 1/68 (2006.01) C07H 21/00 (2006.01) C12N 9/00 (2006.01) C12N 11/00 (2006.01) G01N 33/543 (2006.01)
Patent
CA 1333366
A nucleic acid probe, wherein the label is a catalytically active enzyme, is provided. The nucleic acid of the probe is a DNA or a RNA and comprises a segment of at least 20 nucleotides in the sequence complementary to that of the target segment of the probe. In the probe, the nucleic acid and enzyme are covalently joined by a linker, which is bonded, at one end, to the 5'-carbon of the 5'-nucleotide of the nucleic acid and, at the other end, to the enzyme and which comprises an hydrazone or hydrazide group. Also provided is a nucleic acid, which is an intermediate for making a probe according to the invention and which is derivatized, at the 5'-carbon of the 5'-nucleotide, with a moiety with a free hydrazino group. Further provided are methods of making a nucleic acid probe of the invention. In one such method, a nucleic acid of the invention is reacted with an enzyme, which is derivatized to have an aldehyde group. In another such method, a probe of the invention, with an hydrazone in the linker joining the nucleic acid and enzyme, is reduced with cyanoborohydride; such reduction changes the hydrazone to an hydrazide but otherwise does not substantially affect the probe. Further, a method is provided for detecting a nucleic acid analyte by carrying out a nucleic acid probe hybridization assay with a probe according to the invention for the analyte. Finally, the invention provides the compound 4-N'-benzylamidobenzaldehyde and nucleic acid probes labeled with 2,4-dinitrophenyl or 4-N'-benzylamidophenyl through a linker which comprises an hydrazone or hydrazide group.
610032
Macrae & Co.
Siska Diagnostics Inc.
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