基于实时荧光PCR法定量检测食品中鸡源性成分

Quantitative Detection of Chicken-derived Components in Food Based on Real-time PCR

肉类食品真伪鉴定是目前食品安全检测领域的重点研究内容之一。为准确定量检测食品中的鸡源性成分,根据现行检验检疫标准(SN/T 2727—2010)合成引物及探针,并拓展了特异性、灵敏度、检出限和定量分析等试验。结果显示:特异性试验中,仅鸡成分检测出现特异性扩增,其他动物组织均未出现特异性扩增;梯度稀释鸡源性DNA模板原液进行灵敏度评价,发现引物和探针灵敏度较高,可以检测到100 fg级的鸡组分模板DNA,且建立的标准曲线具有良好的线性关系,R2达0.99以上,定量准确;加工制品中的验证试验结果显示,合成的引物及探针具有良好的适用性和定量检测能力。以上研究结果均说明,此引物和探针适用于市场食品中鸡源性成分的定性、定量检测。本研究为今后TaqMan实时荧光PCR法不同动物源性引物和探针的自主研发奠定了基础。

At present,the identification of meat products is emphatically studied in the field of food safety.In order to establish a quantitative detection method for chicken-derived components in food,primers and probes were synthesized according to current standard of inspection and quarantine(SN/T 2727—2010),and specificity test,sensitivity test,detection limit and quantitative analysis were expanded.The results showed that the specific amplification was found only in chicken-derived components instead of other animal tissues;gradient dilution of chicken-derived DNA template were used to evaluate the sensitivity,and it was found that the sensitivity of primers and probe was high,which could detect the DNA template at 100 fg level,and a good linear relationship was observed in the standard curve,R2 could reach above 0.99 with an accurate quantitative detection.For the processed products,the synthesized primers and probes were with good applicability and quantitative detection capacity.It was concluded that the primers and probes were suitable for qualitative and quantitative detection of chicken-derived components in food markets.A foundation was laid for the independent development of different animal-derived primers and probes of Taq Man real-time PCR in the future.