Abstract
The aim of this work was to use TaqMan Real-Time PCR for quantitative authentication of chicken and turkey meat.To meet this purpose, a specific pair of primers and TaqMan probe was used. The test was aimed at identifying the reaction cycle of turkey and chicken meat using by two sets of primers. With first set of primer designed for chicken we obtained the following results: Cp = 16.18 for 100% chicken DNA Cp = 29, 18 100% turkey DNA It was also amplified DNA of pig that exceeded the detection threshold fluorescence intensities in the 31.07 cycle (Cp = 31.07). Using primers designed for turkey we obtained the following results Cp = 31.16 for 100% CHDNA, Cp =16.18 100% TDNA. It was also amplified the 100% DNA of rabbit in 31.63 cycle (Cp = 31.63) and deer in cycle 32 (Cp = 32). The DNA of all other animal species was amplificated after more than 35 cycles (Cp >35). It follows that the second detection primer pair is specific enough to unrelated species of animals by 30 cycles of the reaction. Species authentication based on DNA analysis from this perspective overcomes all the shortcomings of proteins. At present, DNA analysis use different types of PCR. Is the most progressive Real-time PCR, which is suitable for the specific use of detection (primers and TaqMan probe). The TaqMan Real-time PCR is within the sensitivity and specificity, clearly one of the best methods for identifying the species of chicken and turkey meat. The specificity of this method, however, depends primarily on the specificity of the primers and TaqMan probe. The 30 cycle reaction was chosen by us as the threshold for specificity using primers for authentication chicken and turkey meat.
Highlights
For food producers is attractive for economic reasons to replace expensive components with cheaper.they falsify especially expensive products or products that are produced in large volumes due to higher profits (Lees, 2003; Peris and Escuder-Gilabert, 2009).Popelka et al (2002) the adulteration of food is associated with the deteriorating quality of food.In Slovakia is the necessary verification of genuineness of certain products as a necessary part of a comprehensive examination of quality of goods with regard to consumer protection, together with the fight against counterfeiting of products in the package itself or directly for sales (Takáčová and Bugarský, 2010).At present is more and more used PCR method allowing the direct quantification of PCR products during the amplification reaction – Real-Time PCR
The aim of this work is to evaluate the determination of species specificity of primers for detection of turkey and chicken
The sequence of the primers and TaqMan probes of the first and second sets of detection are listed in Table 1 and Table 2
Summary
For food producers is attractive for economic reasons to replace expensive components with cheaper.they falsify especially expensive products or products that are produced in large volumes due to higher profits (Lees, 2003; Peris and Escuder-Gilabert, 2009).Popelka et al (2002) the adulteration of food is associated with the deteriorating quality of food.In Slovakia is the necessary verification of genuineness of certain products as a necessary part of a comprehensive examination of quality of goods with regard to consumer protection, together with the fight against counterfeiting of products in the package itself or directly for sales (Takáčová and Bugarský, 2010).At present is more and more used PCR method allowing the direct quantification of PCR products during the amplification reaction – Real-Time PCR. The aim of this work is to evaluate the determination of species specificity of primers for detection of turkey and chicken. Assess the specificity of the first and second detection kits examining cross-reaction with other species.
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