Immuno-PCR Assay for Sensitive Detection of Proteins in Real Time

Methods Mol Biol. 2015:1318:139-48. doi: 10.1007/978-1-4939-2742-5_14.

Abstract

The immuno-PCR (IPCR) assay combines the versatility and robustness of immunoassays with the exponential signal amplification power of the polymerase chain reaction (PCR). Typically, IPCR allows a 10-1,000-fold increase in sensitivity over the analogous enzyme-linked immunosorbent assay (ELISA). This is achieved by replacing the signal-producing antibody-enzyme conjugate of an ELISA with an antibody-DNA conjugate that serves as a marker for PCR amplification. The amplification power of the PCR allows for the detection of even single molecules of nucleic acid templates, making it well suited for a broad range of applications. Here, we describe the application of an IPCR assay for detection of trace amount of antigens using ricin as an example.

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antigens / analysis*
  • Biotin / chemistry
  • Buffers
  • DNA / chemistry
  • DNA Restriction Enzymes / chemistry
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Immunoconjugates / chemistry*
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Ricin / analysis*
  • Ricinus / chemistry
  • Sensitivity and Specificity
  • Streptavidin / chemistry
  • Time Factors

Substances

  • Antibodies, Monoclonal
  • Antigens
  • Buffers
  • Immunoconjugates
  • Biotin
  • DNA
  • Ricin
  • Streptavidin
  • DNA Restriction Enzymes