METHOD FOR DETERMINING ACTIVITY OF REVERSE TRANSCRIPTASE

摘要

A method for determining the activity of a nucleotide polymerizing enzyme in a sample, and use of the method for determining HIV 1 RT- and Herpes Simplex DNA-polymerase activity. The enzyme is captured by means of a monoclonal antibody which is immobilized to a solid carrier and is capable of binding the enzyme without detrimentally effecting the enzyme activity. Contaminants and disturbing factors are removed and the nucleotide polymerization starts by the addition of a reaction solution containing a primer/template construct and nucleotides substrate, the reaction conditions being chosen such that they promote permanent association between antibody enzyme- and primer/template constructs. When necessary nucleotide substrate, primer/template and reaction solution are washed away from the newly synthesized polymer, and the amount of nucleotide which has been incorporated into the polymer is determined, and the activity of the enzyme is determined with the guidance of this determination.