The Development of Species-specific Real-time PCR Assays for the Detection of Pheasant and Quail in Food
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The Development of Species-specific Real-time PCR Assays for the Detection of Pheasant and Quail in Food James Chisholm & Ana Sánchez & Joy Brown & Heather Hird
Received: 30 November 2007 / Accepted: 4 March 2008 / Published online: 9 May 2008 # Springer Science + Business Media, LLC 2008
Abstract The development of species-specific real-time PCR assays for the detection of pheasant and quail in commercial food products are reported. Real-time PCR primer and probe sets were designed to detect the mitochondrial cytochrome b gene of pheasant (Phasianus colchicus) and quail (Coturnix coturnix) and were optimized to achieve species specificity. The efficiency and sensitivity of the assays were determined and their applicability to the analysis of commercial samples assessed. The assays successfully detected pheasant and quail in complex food matrices of raw, oven-cooked, and autoclaved meat, demonstrating their suitability for use in enforcement and food control laboratories. Keywords Real-time PCR . Pheasant . Quail . Species . Detection
Introduction Pheasant (Phasianus colchicus) and quail (Coturnix coturnix) are relatively specialized meats that are eaten less frequently than chicken, pork, and beef. They can be found in gourmet foods that tend to be eaten on special occasions J. Chisholm (*) : J. Brown : H. Hird Central Science Laboratory, Sand Hutton, York, North Yorkshire YO41 1LZ, UK e-mail: [email protected] A. Sánchez Instituto de Investigaciones Marinas, 36208 Vigo, Spain
and because of their specialized nature, command high prices, especially quails eggs, which are usually pickled in brine or vinegar. As consumer’s tastes and choices broaden, the demand for less common, game type meats is likely to grow, together with the need for suitable tests to ensure the accurate labeling of these products. Indeed, a study by Pascoal et al. 2004, which surveyed a range of commercial meat products, identified an approximate 50% substitution of mountain quail (Oreortyx pictus) with turkey in a canned quail product, indicating that this type of adulteration and/or substitution is an issue. Many techniques have been applied to allow the enforcement of food labeling regulations, including microscopy, immunochemistry, and molecular biology. Molecular biology-based techniques have been shown to be particularly useful for the identification of meat species in commercial products, and real-time PCR, utilizing TaqMan® technology, is becoming a widely adopted technique, superseding conventional gel-based PCR, which is slower and less sensitive. Studies concerned with the identification of common meat species including chicken, cow, pig, and duck (Dooley et al. 2004; Laube et al. 2003; Matsunaga et al. 1999; Lopez-Andreo et al. 2005; Hird et al. 2005) using real-time PCR technology have been reported recently; however, the number of reports for game meats, for example, pheasant, quail, rabbit, and hare, are relatively limited with only one report of pheasant- and quail-specific assays utilizing conventional gel-based PCR to d
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