食用酱油中曲霉的检测方法  

Detection of Aspergillus spp.in Soy Sauce

作　　者：王建伟[1,2] 刘昌来[2] 周艳红[2] 祭芳[2] 徐剑宏[2] 史建荣[2] 

机构地区：[1]南京农业大学植物保护学院植物病理学系,农业部作物病虫害监测与防控重点开放实验室,江苏南京210095 [2]江苏省农业科学院食品质量安全与检测研究所,农业部食品安全监控重点实验室,江苏省食品安全重点实验室,江苏南京210014

出　　处：《江苏农业学报》2010年第4期848-852,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家科技支撑计划(2006BAK02A28);国家重大科技专项(2009ZX08011-015B);江苏省科技项目(BE2009699;BZ2008119);江苏省农业科技创新项目[CX(08)137]

摘　　要：为了快速检测酱油中黄曲霉毒素和赭曲霉毒素产生菌的污染,利用现代分子技术并结合曲霉菌的一些菌落与孢子特征差异,建立了曲霉菌快速检测方法。结果表明,使用黑曲霉钙调蛋白基因所设计的黑曲霉特异性引物,能够将黑曲霉与其他曲霉分离,黑曲霉中可以扩增出钙调蛋白基因特异性片段,而在米曲霉和黄曲霉中不能扩增到该基因片段。利用aflR等多个已知基因设计特异性引物,扩增结果表明,米曲霉和黄曲霉的菌株并无特异性条带差异。但是,黄曲霉与米曲霉的菌落特征及孢子特征差异显著。由此建立的以PCR检测结合菌株培养特征区分多种曲霉的方法,可以用于检测酱油中黄曲霉毒素和赭曲霉毒素产生菌的污染。In order to identify and evaluate the risk of aflatoxin and ochratoxins in soy sauce,a rapid detection of Aspergillus spp.in soy sauce was established.A pair of primers which amplified the specific fragment from Aspergillus niger,based on the calmodulin gene of Aspergillus niger,was designed,which could differentiate Aspergillus niger from Aspergillus oryzae and Aspergillus flavus.Based on aflR gene,another pair of primers to distinguish Aspergillus oryzae from Aspergillus flavus which produce aflatoxin was designed.The amplification results showed insignificant difference between Aspergillus oryzae and Aspergillus flavus.However,the colony characeristics of Aspergillus flavus and Aspergillus oryzae were clearly different.In conclusion,a method combining PCR detection and colony characteristics in comparison to distinguish Aspergillus species was established,which could be used for evaluation of aflatoxins and ochratoxin in soy sauce.

关 键 词：酱油 曲霉 钙调蛋白基因 aflR基因 PCR 菌落特征 

分 类 号：Q949.327.1[生物学—植物学]