适于肉种掺杂比例实时荧光PCR测定的样品前处理方法  

An Optimized Sample Pretreatment Method for Quantitative Detection of Meat Adulteration by PCR

作　　者：杨君娜[1] 李家鹏[1] 周彤[1] 田寒友[1] 乔晓玲[1] 

机构地区：[1]中国肉类食品综合研究中心,北京100068

出　　处：《肉类研究》2011年第12期25-28,共4页Meat Research

基　　金：北京市科技计划项目(D101110047710003);国家"863"计划项目(2011AA100805)

摘　　要：对适用于肉种掺杂比例实时荧光PCR测定的样品前处理方法及样品均匀性进行研究。正交试验结果表明:稀释倍数为肉、水比1:4(m/V),匀浆机转速12000r/min,匀浆时间8min,此条件下制备的样品均匀性好,样品质量、DNA质量浓度和实时荧光PCR实验Ct值结果都具有较好的均一性,无显著性差异。采用这种样品前处理方法时,肉样的取样量对样品的均匀性没有影响。In this study, an L9（34） orthogonal array design was employed to optimize homogenization conditions for use in the sample pretreatment for quantitative detection of meat adulteration by PCR. Meanwhile, the effect of sample homogeneity on the accuracy of PCR detection was examined using three different sampling amounts： 30, 10 g and 3 g of rump pork. The optimal homogenization conditions were found to be： 1：4 （m/V） meat/water ratio, 12000 r/min homogenizer rotary speed and 8 min homogenization time. Under these conditions, a good homogeneity was obtained for pretreated samples, and the results of determination of sample mass, DNA concentration and real-time PCR Ct also showed a good homogeneity and no significant difference. Further, the optimized homogenization conditions were used to detect pork at the three sampling levels above, with the result that sampling amount had no effect on sample homogeneity.

关 键 词：肉种掺杂比例 实时荧光PCR实验 前处理方法 样品均匀性 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程] O657.3[轻工技术与工程—食品科学与工程]