title: A Short Interspersed Nuclear Element (SINE)-Based Real-Time PCR Approach to Detect and Quantify Porcine Component in Meat Products.
abstract: Real-time PCR amplification of mitochondria gene could not be used for DNA quantification, and that of single copy DNA did not allow an ideal sensitivity. Moreover, cross-reactions among similar species were commonly observed in the published methods amplifying repetitive sequence, which hindered their further application. The purpose of this study was to establish a short interspersed nuclear element (SINE)-based real-time PCR approach having high specificity for species detection that could be used in DNA quantification. After massive screening of candidate Sus scrofa SINEs, one optimal combination of primers and probe was selected, which had no cross-reaction with other common meat species. LOD of the method was 44 fg DNA/reaction. Further, quantification tests showed this approach was practical in DNA estimation without tissue variance. Thus, this study provided a new tool for qualitative detection of porcine component, which could be promising in the QC of meat products.
authors.affiliation: Nanjing University of Chinese Medicine, No. 138 Xianlin Rd, 210023, Nanjing, People's Republic of China.
authors.forename: Chi
authors.initials: C
authors.lastname: Zhang
authors.forename: Xin
authors.initials: X
authors.lastname: Fang
authors.forename: Haopu
authors.initials: H
authors.lastname: Qiu
authors.forename: Ning
authors.initials: N
authors.lastname: Li
mesh_terms: D000818:Animals; D004247:DNA / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D005504:Food Analysis; D006801:Humans; D057230:Limit of Detection; D008461:Meat Products / Q000032:analysis*; D011786:Quality Control; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D020086:Short Interspersed Nucleotide Elements; D013045:Species Specificity; D013552:Swine
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004247:DNA