Development and design of a ‘ready‐to‐use’ reaction plate for a PCR‐based simultaneous detection of animal species used in foods

Abstract

Different TaqMan TM -polymerase chain reaction systems have been developed, which allow the detection of even minute amounts of beef, pork, lamb, goat, chicken, turkey and duck in processed foods. The speciesspecific systems are able to amplify DNA regions with no more than 108 bp in size (exception: duck, 212 bp) located on the single-copy genes cyclic guanosine monophosphate (cyclic GMP) phosphodiesterase, ryanodine receptor and interleukin-2 precursor. The parallel detection of the common ingredient 'meat' produced from mammals and poultry was based on the amplification of a region of the myostatin gene. The limit of detection was determined to be ten genome copies for each system. The relative SD under repeatability condition was below 30%. In addition, a 'ready-to-use' reaction plate has been developed, which makes it possible to investigate the presence of the seven animal species in parallel after a single real-time run.

Quantitative determination of commercially relevant species in foods by real‐time PCR

“…In a mixture prepared of 10% turkey and 90% chicken, 12% turkey was determined using TaqMan PCR. In mixtures containing duck, the genome size along with the difference to the sequence of MYw forward primer (Laube et al. , 2005) has to be taken into consideration.…”

“…Real time PCR using TaqMan TM technology was performed on the Applied Biosystems ABI PRISM TM 7700 Sequence Detection System (Foster City, CA, USA), as described in Laube et al. (2005).…”

“…The copy number (C) was calculated based on DNA concentration determined spectrophotometrically, taking into account the base pair sizes of vector and insert and the genome size described in Gregory (2005). In order to exclude false‐negative results, an internal positive control (Laube et al. , 2005) was used.…”

“…The objective of the present study was to examine the quantitative determination of the animal species cattle, pig, lamb, goat, chicken and turkey based on real‐time PCR systems published in Laube et al. (2005).…”

Advances in Authenticity Testing for Meat Speciation

“…Later on, the same authors used the same target gene and sequencing as a tool for further development of PCR-RFLP methods for meat species differentiation (Girish et al, 2005(Girish et al, , 2007. In fact, sequencing of PCR products has been frequently used as an intermediate step during the development of several methods of PCR-RFLP (Fajardo et al, 2006;Rojas et al, 2009a), species-specific PCR (Haunshi et al, 2009;Santos et al, 2012), and real-time PCR (Laube et al, 2007;Santos et al, 2012;Kane and Hellberg, 2016).…”