The United States of America, As Represented by the Secretary, Department of Health and Human Services

The United States of America As Represented by the Department of Health

Shinji Yamamoto

Walid M Heneine

Thomas M Folks

William M Switzer

Gerardo Garcia-Lerma

Gerardo Garcia Lerma

Assays and kits for the detection of phenotypic resistance of a retrovirus to reverse transcriptase inhibitor-drugs in a biological sample. The assays are based on the direct analysis of the susceptibility of retroviral reverse transcriptase to inhibition by a reverse transcriptase inhibitor drug. The enzymatic activity of the reverse transcriptase is determined by measuring the DNA product produced when an RNA template and a first complementary DNA primer from a suitable region of the encephalomyocarditis virus genome are incubated with a biological sample containing reverse transcriptase in the presence of the drug to which resistance is being determined. The DNA product is amplified and detection of the amplified DNA indicates resistance to the drug employed in the assay. Detection of relatively greater amounts of amplified DNA when certain drugs are used indicates the presence of multiple nucleoside analog resistant strains or mutations.

Method and kit for detecting resistance to antiviral drugs

An assay and kit for the detection of phenotypic resistance of a retrovirus to a reverse transcriptase inhibitor drug in a biological sample. The assay is based on the direct analysis of the susceptibility of retroviral reverse transcriptase to inhibition by a reverse transcriptase inhibitor drug. The enzymatic activity of the reverse transcriptase is determined by measuring the DNA product produced when an RNA template and a first complementary DNA primer from a suitable region of the encephalomyocarditis virus genome are incubated with a biological sample containing reverse transcriptase in the presence of the drug to which resistance is being determined.

The present invention provides a method for detecting the presence of a retrovirus in a biological sample comprising the steps of: a) contacting the biological sample with an RNA template and a complementary DNA primer under conditions whereby the RNA template and the DNA primer will anneal and a DNA strand will be synthesized as an extension from the DNA primer if reverse transcriptase is present in the sample; b) amplifying the synthesized DNA; and c) detecting the amplification of the synthesized DNA, the amplification of the synthesized DNA indicating the presence of reverse transcriptase in the biological sample, thus indicating the presence of a retrovirus in the biological sample.

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Kumamoto, Japan

Shinji Yamamoto