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Comparison of Real-Time PCR Quantification Methods in the Identification of Poultry Species in Meat Products

机译:实时PCR定量方法在肉类产品鉴定中的实时PCR定量方法

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摘要

Poultry meat is consumed worldwide and is prone to food fraud because of large price differences among meat from different poultry species. Precise and sensitive analytical methods are necessary to control poultry meat products. We chose species–specific sequences of the gene to develop two multiplex real-time polymerase chain reaction (real-time PCR) systems: one for chicken ( ), guinea fowl ( ), and pheasant ( ), and one for quail ( ) and turkey ( ). For each species, added meat could be detected down to 0.5 % . No cross reactions were seen. For these two real-time PCR systems, we applied three different quantification methods: (A) with relative standard curves, (B) with matrix-specific multiplication factors, and (C) with an internal DNA reference sequence to normalize and to control inhibition. All three quantification methods had reasonable recovery rates from 43% to 173%. Method B had more accepted recovery rates, i.e., in the range 70–130%, namely 83% compared to 75% for method A or C.
机译:家禽肉在全球消费,并且由于来自不同家禽种类的肉类的价格差异很大,易于食物欺诈。对控制家禽肉类产品需要精确和敏感的分析方法。我们选择基因的种类特异性序列,以开发两种多重实时聚合酶链反应(实时PCR)系统:一个用于鸡肉(),几内亚Fowl()和野鸡(),以及一个用于鹌鹑()和火鸡 ( )。对于每种物种,可以检测到下降至0.5%的添加肉。没有看到交叉反应。对于这两个实时PCR系统,我们应用了三种不同的定量方法:(a)具有相对标准曲线,(b)具有基质特异性繁殖因子,(c)具有内部DNA参考序列以标准化和控制抑制。所有三种量化的方法具有合理的回收率,从43%到173%。方法B具有更高的恢复速率,即70-130%的范围,即53%的方法A或C的75%。

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