Multiplex PCR assay for animal species identification in meat bone meal

Abstract

The authenticity of meat bone meal (MBM) in animal feed is needed to be considered due to inappropriate feed ingredients for certain types of livestock may cause various diseases. This work was aimed to identify animal species in MBM using multiplex PCR assay with 12S rRNA gene as a target region. Total of eight DNA samples were extracted from three species (chicken, bovine, and porcine) which are used as positive controls and three MBM samples. The MBM samples were collected from three different importers. Additionally, a multiplex PCR assay has been performed to identify animal species in MBM. Multiplex PCR of 12S rRNA gene was designed to detect bovine, chicken, and porcine in MBM samples. The PCR products was visualized using 2% agarose gels under the UV light. The results showed that multiplex-PCR of 12S rRNA gene was able to identify bovine which is indicated by 155 bp of DNA band. Chicken and porcine were not found in MBM samples. Meat bone meal samples used in this study might be used to be a feedstuff for poultry, porcine, and other non-ruminants. In conclusion, Multiplex-PCR using mt-DNA 12S rRNA was effective and accurate technique to identify species contained in MBM samples.