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KMID : 1134820170460091097
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Journal of the Korean Society of Food Science and Nutrition
2017 Volume.46 No. 9 p.1097 ~ p.1105
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Use of Real-Time PCR and Internal Standard Addition Method for Identifying Mixed Ratio of Chicken Meat in Sausages
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Lee Nam-Rye
Joo Jae-Young
Yeo Yong-Heon
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Abstract
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This study examined how much chicken meat was in sausage made with pork. Both real-time polymerase chain reaction (PCR) and internal standard addition were used. Fifty ng of chicken DNA was added to the sausages as an internal standard. The addition of standard DNA increased the amplification efficiency of PCR and confirmed the possibility of quantitative analysis. A QIAamp DNA Micro Kit was used to improve the DNA recovery and amplification efficiency. The density of template DNA and primer were suitable for 3.0∼5.0 μL and 0.5 μL, respectively. Each DNA of pig and chicken was diluted in 10-fold from steps 50 ng to 0.05 ng. The detection limit of both pig and chicken meat was more than 0.05 ng and the correlation coefficient of the standard curve was at least 0.98. The result of the quantitative analysis after heat treatment of 3 samples of pigs and chickens mixed at 70:30 showed a 5.7% difference (64.3:35.7) between the expected value and measured value. The quantitative value was changed by affecting the DNA according to the heat treatment (70°C, 10 min). An analysis of the pork and chicken content in sausages showed that it was difficult to detect chicken meat and the quantitative value of DNA according to the Ct value was very low. On the other hand, when adding standard material (50 ng of chicken DNA) to the sausages, the Ct value decreased gradually with increasing chicken mixing ratio. Thus, the mixing ratio of chicken in sausages could be estimated.
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KEYWORD
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gene analysis, sausage, internal standard addition, real-time PCR
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