Appeal No. 95-4404
Application 07/913,121
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 containing reverse
transcriptase and at least one drug known to have reverse transcriptase inhibitory activity
under conditions leading to the production of a cDNA strand complementary to the
template RNA and hybridized thereto if the reverse transcriptase was not inhibited by the
drug;
(C) degrading the RNA template from the RNA-cDNA complex if formed in step
(B) resulting in single stranded cDNA;
(D) hybridizing said cDNA if formed with a chemically modified oligonucleotide
probe or probes to permit capture and/or detection of the cDNA-probe complex formed;
14. A method for detecting and/or quantitating the reverse transcriptase
inhibitory activity of a compound which comprises:
(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 an appropriate reverse
transcriptase standard and at least one compound suspected to be a reverse
transcriptase inhibitor under conditions lead to the product of a cDNA strand
complementary to the template RNA and hybridized thereto if the reverse transcriptase
was not inhibited by the drug;
(C) degrading the RNA template from the RNA-cDNA complex if formed in the
presence of the compound in step (B) resulting in single stranded cDNA;
(D) hybridizing said cDNA if formed in the presence of the compound 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 if formed in the presence of the
compound in step (D) from unreacted probe, wherein steps (D) and (E) can be carried out
sequentially or simultaneously; and
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