PCR Test for Exogenous DNA Detection with Applications in Infectious Disease Diagnosis, Gene Therapy Safety Trials and Gene Doping Surveillance

Technology #13468

This TaqMan-based PCR testing format will allow scientists to quickly, accurately and inexpensively determine the presence of exogenous DNA. It is user-friendly, capable of testing large numbers of samples, and provides a binary readout (a sample is deemed positive or negative for the presence of the target sequences). Employing an internal threshold control (ITC) template, University of Florida researchers simplified PCR testing down to a single reaction. Their improved test overcomes many of the limitations associated with existing assays, reducing the risk of false positives and negatives. Sensitive and specific tests for detecting exogenous DNA molecules are in high demand for infectious disease diagnosis, gene therapy clinical safety trials, and gene doping surveillance programs. This fast, accurate and easy-to-use test is well-positioned in the global PCR market, which is projected to reach $27 billion by 2015.

Application

A sensitive and specific test for detecting exogenous DNA molecules

Advantages

  • Collects data automatically and provides fast, accurate, low-cost results within two to three hours
  • Analyzes several samples at once, permitting faster processing
  • Supports the collection of assay results in centralized databases, facilitating further analysis
  • Lowers the risk of false positives or negatives, ensuring accuracy
  • Compatible with human and animal DNA analysis, broadening the potential market

Technology

Existing Taqman real-time PCR tests use specific sequence probes to detect exogenous DNA. One of the major problems associated with these analyses is the high number of false positive signals (caused by non-targeted exogenous or endogenous DNA sequences) and false negative signals (caused by impurities that inhibit PCR). Although multiplex Taqman PCR assays with extra primer-probe sets have been used to improve accuracy, differences between targets can still lead to different detection efficiencies. Researchers at the University of Florida have developed a single-reaction test that incorporates an internal threshold control (ITC). The target sequence and ITC template are co-amplified by the same primers, but are detected by different probes, each with a unique fluorescent dye.