Methods for the detection of beef and pork in foods using real‐time polymerase chain reaction

Abstract

Summary Two TaqMan™‐polymerase chain reaction (PCR) systems have been developed which permit the detection of even minute amounts of beef and pork in processed foods. In these methods cattle‐specific primers amplified a fragment from the phosphodiesterase gene having a length of 104 base pairs (bp), and the swine‐specific primers amplified a fragment from the ryanodin gene having a length of 108 bp. Beyond this, a third TaqMan™‐PCR system, developed on the basis of the detection of the myostatin gene, permits a reliable exclusion of false‐negative results by detecting meat from a variety of mammals or poultry in the material to be tested.

The fate of DNA of transgenic inulin synthesizing potatoes in pigs

“…1. For all tissue samples a PCR system developed by Laube et al (2003) was selected to determine the integrity and quality of DNA extracted. 2.…”

The Application of PCR-Based Methods in Food Control Agencies – A Review

“…Köppel et al (2009) have for example described the implementation of a heptaplex Realtime PCR assay for the simultaneous identification and quantification of DNA from beef, pork, chicken, turkey, horse meat, sheep (mutton) and goat. Although such multiplex approaches will greatly accelerate meat species identification, results generated must be taken with caution as several meat products are produced with widely varying fat and tissue composition, thus the DNA extractable from similar meat products might vary greatly (Laube et al, 2003).…”

Meat

“…Samples are minced and homogenized and DNA is extracted using a modified cetyltrimethylammonium bromide (CTAB) method (Laube et al, 2003) based on a procedure developed by Tinker et al (1993). One hundred milligrams of homogenate is incubated on a shaker together with 500 mL of CTAB solution (CTAB 20 g/L, NaCl 1.4 mol/L, Tris-HCl 0.1 mol/L, Na 2 -EDTA 20 mmol/L; pH 8.0) and 20 mL of proteinase K (20 mg/mL) at 65 C for 90 min.…”

“…The assay is composed of two elements: general proof of the presence of meat, and the identification of species-specific sequences (Laube et al, 2003). A common meat-specific PCR system has been established detecting mammalian and poultry species on the basis of a region of the myostatin gene (GenBank accession: AF320998 for cattle and AY448007 for chicken).…”

“…The plots generated by real-time PCR show the fluorescence representing the increasing target copy number as a function of cycle number. Using the MYw primer-probe system (Laube et al, 2003), specific for mammals and poultry (Table 7.1), the DNA of all animal species except salmon, as well as human beings, could be amplified and clearly detected. In addition, the MYw primer-probe system was applied to DNA from onion, garlic, paprika, and pepper.…”