摘要
以鸭肉细胞核中基因序列(IL-2)为目标序列,设计特异性引物和TaqMan探针,设计并构建扩增内标,针对内标设计TaqMan探针,通过对聚合酶链式反应(PCR)反应体系和反应条件的优化,建立用于检测食品中鸭肉成分的添加有扩增内标的的荧光PCR方法。通过扩增曲线和Ct值分析该方法的特异性、检测限和灵敏度。结果表明:TaqMan探针实时荧光PCR法具有很高的特异性,所建立的方法能够检测出0.5ng的鸭组分DNA,灵敏度可达0.1%。应用该方法对市售38份样本进行抽样检测,结果令人满意。
Fluorescent real-time PCR system for the assay of duck meat in foods was established on the basis of a pair of primers and the TaqMan probe specific for duck was designed according to the sequence of duck nuclear gene （IL-2）. We also designed and established an internal amplification control （IAC） and TaqMan probe for IAC. Results showed that the specificity of TaqMan real-time PCR was reliable and it could detect the presence of duck meat even when the concentration of DNA was reduced to 0.5 ng. The sensitivity of this method was 0.1%. Its accuracy was verified with 38 food samples from the market.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第24期92-96,共5页
Food Science
基金
国家"863"计划项目(2011AA100 805-1)
农业部公益性行业(农业)科研专项(201303083-2)
国家农业科技成果转化资金项目(2012GB2C100151)
关键词
细胞核基因
扩增内标
实时荧光聚合酶链式反应
鸭肉
nuclear DNA
intemal amplification control
fluorescent real-time polymerase chain reaction
duck meat
