A METHOD FOR MEASURING DNA POLYMERIZATION AND APPLICATIONS OF THE METHOD

摘要

A method for measuring DNA-dependent DNA polymerisation in a biological sample, is described. The method comprises the steps of providing a primer with a single stranded short specific sequence, which is unable to base pair internally, bound to a solid phase; contacting the primer construct with a reaction mixture containing a single stranded deoxynucleotide template with a part of the sequence complementary to the primer and the four deoxynucleoside triphosphates, one of which is modified so that it is specifically recognized by a labeled antibody; adding a biological sample comprising the DNA polymerase, such as retrovirus reverse transcriptase (RT), to the mixture; allowing the polymerase reaction to proceed; incubating the immobilized reaction product with the labeled antibody; detecting the amount of bound labeled antibody; and measuring the amount of incorporated modified deoxynucleoside triphosphate, as a measure of the DNA polymerisation, which may be used drug susceptibility testing. A commercial package is also disclosed.