Patent ID: 6316182
Filing Date: 2001-11-13
Classification: C12Q

Abstract:
A method for for detecting reverse transcriptase in a sample suspected of containing reverse transcriptase comprising the steps of:(A) incubating a synthetic heteropolymeric RNA template molecule containing 40-500 nucleotides with an oligonucleotide primer complementary to a portion of said RNA molecule and of sufficient length to form a stable template-primer complex;(B) contacting the complex formed in step (A) with a sample suspected of containing reverse transcriptase under conditions leading to the production of a cDNA strand complementary to the template RNA and hybridized thereto if reverse transcriptase was present;(C) degrading the RNA template from the RNA-cDNA complex formed in step (B) resulting in single stranded cDNA;(D) hybridizing said cDNA with a chemically modified oligonucleotide probe or probes to permit capture and/or detection of the cDNA-probe complex formed;(E) separating the cDNA-probe complex formed in step (D) from unreacted probe, wherein steps (D) and (E) may be carried out sequentially or simultaneously; and(F) detecting the presence of reverse transcriptase by detecting a label in a labeled cDNA complex, wherein said label is introduced into said complex from labeled oligonucleotide primer, labeled oligonucleotide probe or a labeled third oligonucleotide when the primer is unmodified.