Screening and Validation of a Novel Universal Target DNA Sequence for Simultaneous Identification and Differentiation of Bovine- and Ovine-/caprine- Derived Ingredients in Ruminant Feeds

Abstract Background: Ruminant feeds containing bovine- and ovine-/caprine- derived ingredients would increase the risk of transmissible spongiform encephalopathies (TSE) spread. Results: In this study, a novel universal target DNA sequence was screened through genomic sequence alignment of 53 species, and an efficient and accurate PCR method was further developed to simultaneously identify and distinguish bovine- (125 bp) and ovine-/caprine- (107 bp) derived ingredients in ruminant feeds. The specificity of our PCR method was tested with 20 animal DNA and 3 feed material DNA. This novel universal target DNA sequence was specifically detected in only bovine species (including cattle, buffalo, yak, and zebu) and ovine/caprine species (including sheep and goat). Sensitivity analysis demonstrated that through this universal target sequence, as low as 1% heat-treated (133 ℃, 300 kPa for 20 min) meat and bone meal (MBM) of bovine or ovine species could be detected from in ruminant feeds. Conclusions: In conclusion, this study screened a novel universal target DNA sequence, designed a universal primer pair, and developed an efficient and accurate PCR method for the simultaneous identification and differentiation of bovine- and ovine/caprine- derived ingredients in ruminant feeds.