title: Development of qualitative and quantitative PCR analysis for meat adulteration from RNA samples.
abstract: Total RNA samples were used to establish qualitative and quantitative PCR-based methods for assessing meat adulteration. The primers were designed based on the mRNA sequences of troponin I (TnI), mitochondrial ribosomal protein (MRP) and tropomodulin genes to distinguish chicken, pork, goat, beef and ostrich. There was no cross reaction between the primers, and the detection limit of the cDNA template was 0.01 and 20 ng in simplex PCR and multiplex PCR, respectively. In the low temperature storage test, the detection limits of cDNA template with 10 and 1 ng were determined at 4 °C and -80 °C. In quantitative assay, the precision of real-time PCR analysis expressed as a coefficient of variation (CV) ranged from 0.25% to 5.24% and the trueness, expressed as an error, ranged from 0.28% to 6.98% for adulteration. Thus, herein, we provided alternative tools for the assessment of meat adulteration using mRNA-based PCR methods.
authors: Center for Shockwave Medicine and Tissue Engineering, Department of Medical Research, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan. Electronic address: cjaiho@yahoo.com.tw.
authors: Jai-Hong
authors: JH
authors: Cheng
authors: Department of Food Science, National Pingtung University of Science and Technology, Pingtung, Taiwan. Electronic address: m9436016@mail.npust.edu.tw.
authors: Hsiao-Ting
authors: HT
authors: Chou
authors: Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung, Taiwan. Electronic address: leemengshiou@mail.cmu.edu.tw.
authors: Meng-Shiou
authors: MS
authors: Lee
authors: Department of Food Science, National Pingtung University of Science and Technology, Pingtung, Taiwan. Electronic address: ssheu@mail.npust.edu.tw.
authors: Shyang-Chwen
authors: SC
authors: Sheu
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics; D002645:Chickens / Q000235:genetics; D017931:DNA Primers; D005506:Food Contamination / Q000032:analysis*; D061353:Food Storage; D006041:Goats / Q000235:genetics; D008460:Meat / Q000032:analysis* / Q000592:standards; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D020310:Struthioniformes / Q000235:genetics; D013552:Swine / Q000235:genetics
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger
keywords: Meat adulteration; Mitochondrial ribosomal protein; PCR; Tropomodulin; Troponin I; mRNA