婴儿配方粉中阪崎肠杆菌荧光定量PCR检测方法的建立  

Fluorescent quantitative PCR method for detecting Enterobacter sakazakii in infant powder

作　　者：卢雁[1] 王淼[1] 房玉国[1] 王旻[1] 王宇[1] 赵长利[1] 

机构地区：[1]国家乳制品质量监督检验中心,哈尔滨150086

出　　处：《中国乳品工业》2008年第7期47-49,56,共4页China Dairy Industry

摘　　要：建立奶粉中阪崎肠杆菌荧光定量PCR检测方法,为准确、快速地定量检测阪崎肠杆菌奠定基础。根据阪崎肠杆菌16S rDNA基因序列,设计特异引物和荧光标记探针并合成,用PCR扩增产物克隆的重组质粒作为阳性对照,经紫外分光光度计测定重组质粒浓度,梯度稀释后作为标准品绘制标准曲线,其Ct值与模板浓度有良好的线形关系,相关系数R2值为:0.997,可以用于阪崎肠杆菌浓度鉴定。用该方法检测阪崎肠杆菌样品21份,阳性4份,阳性检出率为19%,与常规检测方法结果完全符合。本文建立的检测阪崎肠杆菌的荧光定量PCR方法,较常规技术更为简便、快速,有很好的应用前景和研究价值。To establish fluorescent quantitative PCR method for detecting Enterobacter sakazakii （E. sakazakii） in powder. According to the specific sequence of E. sakazakii 16s rDNA genes, the primers and the fluorescent probe were designed and synthesized. The positive recombinant plasmid would be used as standard quantitative template to make the standard curve for sample detection. The standard curve indicated the linear relationship between Ct and template concentration, R^2=0.997. The detection of 21 cases of the infant powder, isolated of E. sakazakii were found in 4 samples（19%）.In conclusion, the fluorescent Quantitative PCR method for the detection of E. sakazakii is simple and accurate, has a good applying perspective and research worth.

关 键 词：阪崎肠杆菌 荧光定量 阈值 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]