Sul Suyeon, Kim Mi-Ju, Lee Jung-Min, Kim Sung-Yeon, Kim Hae-Yeong
Institute of Life Sciences and Resources, Department of Food Science and Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea.
J Food Prot. 2020 Jun 1;83(6):984-990. doi: 10.4315/JFP-19-583.
In this study, we developed a rapid on-site detection method by using direct ultrafast PCR coupled with a microfluidic chip to identify the presence of chicken meat in processed ground meat products. Chicken-specific PCR primer targeting mitochondrial 16S rRNA gene was newly designed, and its specificity was confirmed against 17 other animal species and 4 different chicken meat samples from different countries of origin. The sensitivity of the chicken-specific ultrafast PCR was 0.1 pg of chicken DNA. To evaluate the limit of detection of the direct ultrafast PCR method, different percentages of chicken meat mixed with pork or beef were prepared. The limit of detection of the direct ultrafast PCR method for the chicken meat-pork and chicken meat-beef mixtures was 0.1% for both raw meat and autoclaved meat. This method was used for 15 commercialized processed ground meat products. In this method, the target sequence was successfully amplified, and the presence of chicken meat in processed ground meat products was identified within approximately 25 min, including the time for sample preparation. Thus, our study shows that this developed direct ultrafast PCR method is a rapid and accurate method for on-site detection of chicken DNA in commercial food products.
在本研究中，我们开发了一种快速现场检测方法，即使用直接超快聚合酶链反应（PCR）结合微流控芯片来鉴定加工碎肉产品中鸡肉的存在情况。新设计了针对线粒体16S rRNA基因的鸡特异性PCR引物，并针对17种其他动物物种以及来自不同原产国的4种不同鸡肉样品确认了其特异性。鸡特异性超快PCR的灵敏度为0.1 pg鸡DNA。为了评估直接超快PCR方法的检测限，制备了不同比例的鸡肉与猪肉或牛肉混合的样品。对于生肉和高压灭菌肉，直接超快PCR方法对鸡肉-猪肉和鸡肉-牛肉混合物的检测限均为0.1%。该方法用于15种商业化加工碎肉产品。在此方法中，目标序列成功扩增，并且在包括样品制备时间在内的约25分钟内鉴定出加工碎肉产品中鸡肉的存在情况。因此，我们的研究表明，这种开发的直接超快PCR方法是一种快速、准确的现场检测商业食品中鸡DNA的方法。
