Real-time PCR detection of ruminant DNA.
To control the spread of bovine spongiform encephalopathy, several DNA methods have been described for the detection of the species origin of meat and bone meal. Most of these methods are based on the amplification of a mitochondrial DNA segment. We have developed a semiquantitative method based on real-time PCR for detection of ruminant DNA, targeting an 88-bp segment of the ruminant short interspersed nuclear element Bov-A2. This method is specific for ruminants and is able to detect as little as 10 fg of bovine DNA. Autoclaving decreased the amount of detectable DNA, but positive signals were observed in feeding stuff containing 10% bovine material if this had not been rendered in accordance with the regulations, i.e., heated at 134 degrees C for 3 instead of 20 min.
Mendoza-Romero
Luis
L
Labocor, S. L. Poligono Industrial La Mina Nave, 38 Colmenar Viejo, Madrid, Spain.
Verkaar
Edward L C
EL
Savelkoul
Paul H
PH
Catsburg
Arnold
A
Aarts
Henk J M
HJ
Buntjer
Jaap B
JB
Lenstra
Johannes A
JA
D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002417:Cattle; D004272:DNA, Mitochondrial / Q000032:analysis*; D016643:Encephalopathy, Bovine Spongiform / Q000517:prevention & control* / Q000635:transmission; D005506:Food Contamination; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012418:Ruminants; D012680:Sensitivity and Specificity; D020086:Short Interspersed Nucleotide Elements / Q000235:genetics*; D013045:Species Specificity
D004272:DNA, Mitochondrial