title: Direct-multiplex PCR assay for meat species identification in food products.
abstract: This is the first time that direct PCR - DNA amplification without prior DNA extraction - was successfully developed and fully validated for rapid and economical simultaneous identification of six commonly consumed meat species. To achieve this, six species-specific primers were selected from previous reports and newly designed from the mitochondrial cytochrome b (cyt b), cytochrome oxidase I (COI), and 12s rRNA gene. The assay generated PCR products of 100, 119, 133, 155, 253, and 311 bp for pork, lamb/mutton, chicken, ostrich meat, horsemeat and beef, respectively. Validation showed that the assay is robust, rapid, economical, reproducible, specific, and sensitive down to 12,500 mitochondrial copy (equating to seven fg). It could be used with a variety of raw meats and products, including highly degraded and processed food samples. This proposed method will be greatly beneficial to the consumers, food industry, and law enforcement.
authors.affiliation: Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, 90112, Thailand.
authors.forename: Thitika
authors.initials: T
authors.lastname: Kitpipit
authors.affiliation: Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, 90112, Thailand.
authors.forename: Kuangtiwa
authors.initials: K
authors.lastname: Sittichan
authors.affiliation: Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, 90112, Thailand. Electronic address: pthanakiatkrai@gmail.com.
authors.forename: Phuvadol
authors.initials: P
authors.lastname: Thanakiatkrai
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics; D002645:Chickens / Q000235:genetics; D017931:DNA Primers; D008460:Meat / Q000032:analysis*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D012756:Sheep / Q000235:genetics; D013045:Species Specificity; D013552:Swine / Q000235:genetics
chemical_list: D017931:DNA Primers; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S
keywords: Direct PCR; Identification; Meat species; Multiplex PCR; Validation