| 摘要 |
The present invention concerns a method to detect the presence of an enzyme in in vivo or in vitro tissue or cell, which method comprises:(a) obtaining a tissue or cell sample to be analyzed;(b) contacting the tissue or cell sample with a substrate of the structure selected from the group consisting of:X=H or one or more natural or synthetic amino acids with or without amino blocking groups,Y=H or one or more natural or synthetic amino acids with or without amino blocking groups,wherein X and Y are the same or different and are amino acid sequences of between about 1 to 1,000,000 amino acids wherein each amino acid is the same or a different amino acid, with the proviso that at least one of X or Y is at least one amino acid;(c) when an enzyme is present in the tissue or cell sample which degrades X, Y and combinations thereof, fluorescent cresyl violet is released in the tissue sample producing a color change;(d) detecting the presence and amount of the enzyme present by the detection and quantification of the fluorescence produced; and(e) optionally comparing the fluorescence to a pre-calibrated fluorescence scale to quantify the fluorescence present. A diagnostic kit for use and a method to prepare amino acid cresyl violet derivatives are described.
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