A Short Interspersed Nuclear Element (SINE)-Based Real-Time PCR Approach to Detect and Quantify Porcine Component in Meat Products

• DOI: 10.5740/jaoacint.15-056
• Publication Date: 2015-10-27T01:54:39Z
• Authors: Chi Zhang; Xin Fang; Haopu Qiu; Ning Li
• Keywords: Quality Control; Swine; DNA; Real-Time Polymerase Chain Reaction; 01 natural sciences; Meat Products; Species Specificity; Limit of Detection; Animals; Humans; DNA Probes; Food Analysis; DNA Primers; Short Interspersed Nucleotide Elements; 0105 earth and related environmental sciences

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.