소, 돼지, 가금육류의 신속한 동정을 위한 TaqManⓇprobe를 이용한 real-time PCR 개발

Abstract

Species-specific <TEX>$TaqMan^{(R)}$</TEX> probe-based real-time PCR assays were developed for detection of beef, pork, chicken, duck, goose and turkey. The primer and probe sets used in this study were designed to be complementary to fibroblast growth factor (FGF) for cattle and pig, mitochondrial NADH dehydrogenase (ND) subunit 3 and ND2 for chicken and duck, 12S rRNA for goose and turkey, respectively. As internal positive control we used conserved region in the ribosomal 18S RNA gene to ensure the accuracy of the detection of target DNA by real-time PCR. We confirmed that real-time PCR assays with the primer and probe sets were positive for cattle, pig and chicken intended target animal species with no cross-reactivity with other non-target animal species. Only >50 ng DNA of beef show cross-reactivity in the determination of duck. Using species-specific primer and probe sets, it was possible to detect amounts of 0.1 ng DNA of cattle and pig, 1.0 pg DNA of chicken, duck and turkey, and 0.1 pg DNA of goose for raw samples, respectively. The detection limits were 0.1 ng DNA of cattle, 1.0 ng DNA of pig and 1.0 pg DNA of chicken for DNA mixtures (beef, pork and chicken) extracted from heat-treated (<TEX>$121^{\circ}C$</TEX>/5 min) meat samples. In conclusion, it can be suggested that the <TEX>$TaqMan^{(R)}$</TEX> probe-based assay developed in this study might be a rapid and specific method for the identification of meat species in raw or cooked meat products.