Sensitive Determination of Infectious Titer of Recombinant Adeno-Associated Viruses (rAAVs) Using TCID50 End-Point Dilution and Quantitative Polymerase Chain Reaction (qPCR)

Cold Spring Harb Protoc. 2020 May 1;2020(5):095653. doi: 10.1101/pdb.prot095653.

Abstract

Adeno-associated virus (AAV) recombinants are currently the vector of choice for many gene therapy applications. As experimental therapies progress to clinical trials, the need to characterize recombinant adeno-associated viruses (rAAVs) accurately and reproducibly increases. Accurate determination of rAAV infectious titer is important for determining the activity of each vector lot and for ensuring lot-to-lot consistency. The following protocol developed in our laboratory uses a 96-well TCID50 format and quantitative polymerase chain reaction (qPCR) detection for the determination of rAAV infectious titer.

MeSH terms

  • Algorithms
  • DNA, Viral / genetics*
  • Dependovirus / genetics*
  • Dependovirus / isolation & purification
  • Genetic Vectors / genetics*
  • Genetic Vectors / isolation & purification
  • Genome, Viral / genetics*
  • HeLa Cells
  • Humans
  • Real-Time Polymerase Chain Reaction / methods*
  • Recombination, Genetic
  • Reproducibility of Results

Substances

  • DNA, Viral