摘要
大豆粕、芝麻粕、花生壳和板栗壳是常见的几种可可粉外源掺假成分,根据它们所特有的基因片段设计引物,以高等植物18SrDNA基因为内参照基因,应用改良CTAB法抽提材料DNA,对分离的DNA进行PCR扩增并分析,建立了可可粉中这几种外源植物成分的快速检测方法。用建立的PCR方法对可可基因组进行扩增,均无特异条带;扩增出的PCR产物测序结果表明与目标片段一致;将各材料DNA倍比稀释,分别对各个稀释度的DNA进行PCR扩增。结果表明,该方法对大豆粕、芝麻粕、花生壳和板栗壳DNA检测的敏感度分别可达82.5pg/μl、32.5pg/μl、124pg/μl和157pg/μl。模拟样品试验发现,该方法的最低检测限(w/w)分别达0.5%、0.5%、5%和5%,可作为可可粉及其制品中这几种外源成分鉴别检测的有效方法。
A rapid PCR assay for detection of several common exogenous vegetative components,including soybean meal,sesame meal,peanut shell and chestnut shell contained in cocoa powder was described. Species-specific PCR primers were designed and used in this purpose. DNA was extracted by modified CTAB method and confirmed by using 18S rDNA gene as internal control. The specific sequences could be amplified with the templates of the corresponding DNA solutions but not the cocoa genome DNA. The sequences of amplicons were consistent with GenBank sequences in a high degree. This method can detect DNA extracted from soybean meal,sesame meal,peanut shell and chestnut shell each in less than 82.Spg/μl,32.Spg/μl,124pg/μl and 157pg/μl. Also,it can detect corresponding materials less than 0.5%（w/w）,0.5%（w/w）,5%（w/w）and 5%（w/w） in cocoa powder. It could be an effective method for detection exogenous vegetative components in cocoa powder.
出处
《生物技术通报》
CAS
CSCD
2008年第5期118-121,共4页
Biotechnology Bulletin
基金
国家质检总局科研项目(2006IK142)
关键词
可可粉
植物源性成分
PCR检测
Cocoa Powder Exogenous vegetative components PCR detection
