摘要 |
<p>A reagent comprising an enzyme-specific cleavable polynucleotide (30, 31) substrate bearing quenched fluorescent moieties (32, 33) is provided, as is a method of making the same. The polynucleotide (30, 31) includes at least one fluorescent moiety (32) sufficiently close to another fluorescent moiety (33) to essentially quench fluorescence of the moieties (32, 33), wherein the fluorescent moieties (32, 33) become readily detectable by fluorometric techniques upon separation by cleaving the polynucleotide. A biological assay method is also provided wherein the reagent is combined with a test sample potentially containing the enzyme (36) being assayed wherein the enzyme (36) will cleave the polynucleotide to release the fluorescent moieties (32, 33), and produce an increase in fluorescence intensity. The assay method finds use in detection and identification of microorganisms, sterilization assurance, pharmaceutical discovery, enzyme assays, immunoassays, and other biological assays.</p> |