PCR identification of ruminant tissue in raw and heat-treated meat meals.
To control the spread of bovine spongiform encephalopathy in cattle through contaminated animal feedstuffs, screening of feed products is essential. We designed five pairs of primers to identify specifically raw and heat-treated tissue from cattle, sheep, goat, deer, and ruminants in general. A forward common primer was designed based on a conserved DNA sequence in the mitochondrial 12S rRNA-tRNA(val)-16S rRNA gene, and reverse primers were designed to hybridize with a species-specific DNA sequence for each species considered. All primers were developed to create a specific PCR product small enough (less than 200 bp) to be suitable for heat-treated material. To evaluate the effect of heat treatment, a severe sterilization condition (133 degrees C at 300 kPa for 20 min) was chosen. Species-specific amplicons were obtained from all types of heat-treated meat meals. Analysis of laboratory-contaminated vegetable meals revealed that the detection limit of the assay was 0.05% for each species analyzed. This PCR-based analysis can be used as a routine method for detecting banned animal-derived ingredients in raw and heat-treated feedstuffs.
Ha
Jeong Chul
JC
School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, San 56-1, Sillim-dong, Gwanak-gu, Seoul 151-921, Korea. hjc1010@hanmail.net
Jung
Wan Tae
WT
Nam
Yong Suk
YS
Moon
Tae Wha
TW
D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis*; D003670:Deer; D016643:Encephalopathy, Bovine Spongiform / Q000635:transmission*; D005506:Food Contamination / Q000032:analysis*; D006041:Goats; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D004272:DNA, Mitochondrial; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; C075526:RNA, ribosomal, 12S; D004247:DNA