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 forw...

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Bibliographic Details
Published in:Journal of food protection 2006-09, Vol.69 (9), p.2241-2247
Main Authors: Ha, J.C, Jung, W.T, Nam, Y.S, Moon, T.W
Format: Article
Language:English
Subjects:
adulterated products
Animal Feed - analysis
animal tissues
animal-based foods
Animals
Base Sequence
beef
Biological and medical sciences
bovine spongiform encephalopathy
Cattle
Deer
detection
DNA - analysis
DNA, Mitochondrial - analysis
Encephalopathy, Bovine Spongiform - transmission
feed contamination
feeds
Food Contamination - analysis
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
Goats
heat treatment
Hot Temperature
Humans
meat meal
meat products
mitochondrial DNA
Molecular Sequence Data
polymerase chain reaction
Polymerase Chain Reaction - methods
prions
product authenticity
risk reduction
RNA, Ribosomal - analysis
RNA, Ribosomal, 16S - analysis
Ruminantia
Sensitivity and Specificity
Sheep
Species Specificity
Swine
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Summary: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°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.
ISSN:0362-028X
1944-9097
DOI:10.4315/0362-028X-69.9.2241