CoRT-PCR assay for the discrimination of endogenous reverse transcriptase activity in eukaryotic cell lines from infectious retrovirus contamination
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United States of America Patent
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Oct 15, 2002
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app pub date -
Jun 14, 2000
filing date -
Jun 14, 1999
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Abstract
The present invention relates to the detection of the presence of reverse transcriptase enzymatic activity in eukaryotic cell lines. Specifically, the present invention comprises a method for discriminating between endogenous reverse transcriptase activity in a substrate cell line (e.g., avian) used as substrates for the manufacture of biological products (including viral vaccines) from exogenous infectious retrovirus by using a combination of transmissibility assay (viral replication) and PBRT assay (RT-PCR). Appropriate levels of viral amplification and number of PCR cycles are determined and employed that permit detection of exogenous infectious RT activity but not endogenous RT activity.
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