[Multiplex fluorescent real-time PCR detection of bovine, goat and sheep derived materials in animal products].
We designed the specific primers and TaqMan probes targeting cytochrome b genes of mitochondrial DNA from bovine, goat and sheep. We used different fluorescents to label the probes. After optimization of reaction conditions, we set up a multiplex fluorescent real-time PCR method to detect bovine, goat and sheep derived materials, simultaneously. We finished the detection tests of 17 kinds of animal DNA and 200 DNA samples from different sources with the developed method and the National Standard GB/T 20190Y-2006 routine PCR method. The coincidence rate of these two methods was 100%. Without electrophoresis or restriction digestion, the developed method could reduce the test time to one third as routine PCR and identify three kinds of animal derived materials including bovine, goat and sheep in one reaction. The developed method was approximately 10 times more sensitive than routine PCR, and was applicable to identifications of bovine, goat and sheep derived materials in feed stuff, meat, milk, pelt and grease, etc. The study showed that the developed real-time PCR method is a rapid, sensitive and efficacious detection assay for bovine, goat and sheep derived materials in animal products.
Shenzhen Entry-exit Inspection & Quarantine Bureau, Shenzhen 518045, China. zengshaoling@gmail.com
Shaoling
S
Zeng
Zhifeng
Z
Qin
Zhouxi
Z
Ruan
Qunyi
Q
Hua
Tikang
T
Lu
Jianqiang
J
Lü
Shukun
S
Chen
Chenfu
C
Cao
Caihong
C
Zhang
Jie
J
Sun
Bing
B
Chen
Shaojing
S
Wu
D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002417:Cattle; D045303:Cytochromes b / Q000235:genetics*; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics; D005506:Food Contamination / Q000032:analysis*; D050259:Genes, Mitochondrial; D006041:Goats; D008460:Meat / Q000032:analysis*; D008931:Mitochondria, Muscle / Q000201:enzymology / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012756:Sheep
D017931:DNA Primers; D004272:DNA, Mitochondrial; D045303:Cytochromes b