A Multiplex-PCR Method for Detection of Bovine Materials in Foodstuffs

摘要

Rapid identification of bovine materials in animal foodstuffs is essential for effective control of a potential source of bovine spongiform encephalopathy (BSE). Here, we have developed a convenient polymerase chain reaction (PCR)-based assay which allowed detection and identification of a bovine-specific genomic DNA sequence in foodstuffs. Simultaneously the assay could assess the DNA quality of the experiment system by amplification of a highly conserved eucaryotic DNA region of the 18-S ribosomal gene, helped to check the reliability of the test result. The amplified bovine specific PCR product was a genomic DNA fragment of lactoferrin, a low copy gene which was different from commonly used bovine specific mitochondria sequence for identification of bovine materials. The specificity of this method was confirmed by the absence of detectable homologous PCR product using reference foodstuffs samples lacking bovine-derived meat and bonemeals, or genomic DNA samples from vertebrates whose offals are commonly included in animal feeds. This method was able to detect the presence of bovine material in foodstuffs when the samples contained more than 0.02% of bovine-derived meat and bone meal. The specificity, convenience and sensitivity of this method suggest that it can be used for the routine detection of bovine-derived materials.