Rahman Md Mahfujur, Hamid Sharifah Bee Abd, Basirun Wan Jefrey, Bhassu Subha, Rashid Nur Raifana Abdul, Mustafa Shuhaimi, Mohd Desa Mohd Nasir, Ali Md Eaqub
a Nanotechnology and Catalysis Research Centre , University of Malayaob , Kuala Lumpur , Malaysia.
b Department of Chemistry , University of Malaya , Kuala Lumpur , Malaysia.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2016;33(1):10-8. doi: 10.1080/19440049.2015.1104558. Epub 2015 Nov 2.
This paper describes a short-amplicon-based TaqMan probe quantitative real-time PCR (qPCR) assay for the quantitative detection of canine meat in chicken nuggets, which are very popular across the world, including Malaysia. The assay targeted a 100-bp fragment of canine cytb gene using a canine-specific primer and TaqMan probe. Specificity against 10 different animals and plants species demonstrated threshold cycles (Ct) of 16.13 ± 0.12 to 16.25 ± 0.23 for canine DNA and negative results for the others in a 40-cycle reaction. The assay was tested for the quantification of up to 0.01% canine meat in deliberately spiked chicken nuggets with 99.7% PCR efficiency and 0.995 correlation coefficient. The analysis of the actual and qPCR predicted values showed a high recovery rate (from 87% ± 28% to 112% ± 19%) with a linear regression close to unity (R(2) = 0.999). Finally, samples of three halal-branded commercial chicken nuggets collected from different Malaysian outlets were screened for canine meat, but no contamination was demonstrated.
本文描述了一种基于短扩增子的TaqMan探针定量实时PCR（qPCR）检测方法，用于定量检测鸡肉块中的狗肉，鸡肉块在包括马来西亚在内的全球各地都很受欢迎。该检测方法使用犬特异性引物和TaqMan探针靶向犬细胞色素b基因的100bp片段。对10种不同动植物物种的特异性检测表明，在40个循环反应中，犬DNA的阈值循环数（Ct）为16.13±0.12至16.25±0.23，其他物种的结果为阴性。该检测方法在故意添加狗肉的鸡肉块中对高达0.01%的狗肉进行了定量测试，PCR效率为99.7%，相关系数为0.995。实际值与qPCR预测值的分析显示回收率很高（从87%±28%到112%±19%），线性回归接近1（R² = 0.999）。最后，对从马来西亚不同销售点收集的三个清真品牌商业鸡肉块样本进行了狗肉筛查，但未发现污染。
