Measuring viral reverse transcriptase activity
OwnerRobert B. Harris, Thomas R. Reynoldsinventors
Life of this patent
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Abstract
In clinical settings as well as in a drug-discovery context, the impact of an agent that may affect reverse transcriptase (RT) can be measured and even quantified by bringing a sample, which may contain an RT, into contact with an RNA template, a primer complementary to the RNA template, and appropriate oligonucleotide-specific primers, under conditions such that they react, in the presence of RT, to form a cDNA product in inverse proportion to the effect of the agent. The amount of any resultant cDNA product then can be measured. The approach is readily implemented as a real-time, quantitative kinetic assay for RT activity.
Claims14
A method for measuring the effect on reverse transcriptase of an agent affecting a sample that may contain a reverse transcriptase, comprising
- (a) bringing said sample into contact with template RNA, a primer complementary to said template RNA, and a plurality of oligonucleotide-specific primers under conditions such that they react, in the presence of a reverse transcriptase, to form a cDNA product in inverse proportion to the effect of said agent, and
- (b) measuring the amount of said cDNA product.
A method for measuring reverse transcriptase in a sample that may contain an HIV reverse transcriptase, comprising
- (a) bringing said sample into contact with template RNA, a primer complementary to said template RNA, and a plurality of oligonucleotide-specific primers under conditions such that they react, in the presence of a reverse transcriptase, to form a cDNA product, wherein said conditions include a reaction temperature less than about the deactivation temperature of said reverse transcriptase, and
- (b) detecting the presence of cDNA product.
Dependencies are inferred from the claim text.