Real-time polymerase chain reaction method for detecting contamination of beef by material from genetically engineered cattle.

Prion protein knockout (PRNP(-/-)) cattle have been developed and may be used to produce bovine material such as serum, collagen, and gelatin. However, genetically engineered animals (GE animals) must not be imported or made commercially available in Japan, because they are not authorized for food use in Japan. We used real-time polymerase chain reaction (real-time PCR) to develop method of detection for neomycin- and the puromycin-resistance genes in beef samples. Plasmids containing the neomycin-resistance gene and the puromycin-resistance gene were used as standard reference molecules. The results clearly showed that the method we developed is capable of quantitatively detecting the neomycin- and the puromycin-resistance genes in the plasmids in the presence of genomic DNA extracted from a beef sample. We also applied the method to testing of beef samples imported from the United States (U.S.). This method will make it possible to monitor beef for contamination by material from GE cattle to assure food safety.