Appeal No. 95-4404
Application 07/913,121
BACKGROUND
Retrovirus particles contain active reverse transcriptase which is essential for
replication of the retroviral single stranded RNA genome. Reverse transcriptase is a
multifunctional enzyme with three distinct enzymatic activities: RNA-dependent DNA
polymerase activity, RNase H activity, and DNA-dependent DNA polymerase activity.
During replication in a host cell, RNA-dependent DNA polymerase activity generates a
DNA strand (cDNA) complementary to the viral RNA and the original viral RNA strand is
degraded by Rnase H activity. Next, a second DNA strand, complementary to the cDNA,
is generated by DNA-dependent DNA polymerase activity. The resultant double stranded
DNA is integrated into the host genome, and the cellular machinery of the host is
commandeered to replicate more retrovirus particles. Retroviruses can be indirectly
detected through detection of reverse transcriptase RNA-dependent DNA polymerase
activity. See the Specification, pages 1 through 3.
DISCUSSION
The claimed invention is directed to a method for detecting reverse transcriptase in
a sample through the ability of the reverse transcriptase to synthesize cDNA. Claims 1-7
comprise: incubating a synthetic heteropolymeric RNA template with a complementary
oligonucleotide primer to form a template-primer complex; incubating the template-primer
complex with the sample to form a cDNA-RNA duplex if reverse transcriptase is present;
-6-
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