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AssigneeThe Trustees of the University of Pennsylvania
InventorsGuangping Gao, James M Wilson, Mauricio R. Alvira
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A method for detecting and isolating AAV sequences in a sample of DNA obtained from tissue or cells is provided, which sample contains DNA and proviral AAV. The method involves subjecting the sample containing DNA to amplification via polymerase chain reaction (PCR) using a first set of primers which specifically amplify a first AAV region. The first AAV region is characterized by having at least 250 nucleotides of AAV capsid nucleic acid sequence, a variable sequence flanked by a sequence of at least 18 nucleotides at the 5′ end of the first AAV region and a sequence of at least 18 nucleotides at the 3′ end of the first AAV region. Each of the 5′ and 3′ at least 18 nucleotides is the same over at least 9 consecutive nucleotides relative to corresponding sequences in an alignment of at least two AAV serotypes. Each of the sets of primers consist of a 5′ primer and a 3′ primer. The method is further useful for identifying AAV sequences in the sample by the presence of amplified proviral AAV sequences.
A method for detecting adeno-associated virus (AAV) nucleotide sequences from an AAV signature region within an AAV capsid gene in a sample from a human or non-human primate, comprising performing a plurality of polymerase chain reactions (PCR) on the sample under conditions that allow for specific amplification of the AAV signature region with a pair of PCR primers consisting of a first PCR primer and a second PCR primer that are designed to amplify an AAV signature region located within the AAV capsid gene in the position of nucleotides 2886 to 3143 of SEQ ID NO:6, wherein the first PCR primer and the second PCR primer form a duplex with AAV sequences which flank said AAV signature region;
Dependencies are inferred from the claim text.