基于基因探针扫描技术高通量检测乳制品中赭曲霉毒素A产生菌  

High-Throughput Detection of Ochratoxin A-Producing Fungi in Dairy Products Using Gene Probe Scanning Technology

作　　者：孔文平 李裕 郭锦材 李灿 朱湘成 陈同强 KONG Wenping;LI Yu;GUO Jincai;LI Can;ZHU Xiangcheng;CHEN Tongqiang(Xiangya International Institute of Translational Medicine,Central South University,Changsha 410083,China;Changsha Stomatological Hospital,Changsha 410006,China;Hunan Provincial Key Laboratory of Food Safety Monitoring and Early Warning,Changsha 410111,China;Hunan Institute of Food Quality Supervision Inspection and Research,Changsha 410000,China;Hunan Engineering Research Center of Combinatorial Biosynthesis and Natural Products Medicine,Changsha 410011,China)

机构地区：[1]中南大学湘雅国际转化医学联合研究院,湖南长沙410083 [2]长沙市口腔医院,湖南长沙410006 [3]食品安全监测与预警湖南省重点实验室,湖南长沙410111 [4]湖南省食品质量监督检验研究院,湖南长沙410000 [5]组合生物合成与天然产物药物湖南省工程研究中心,湖南长沙410011

出　　处：《乳业科学与技术》2022年第1期20-25,共6页JOURNAL OF DAIRY SCIENCE AND TECHNOLOGY

基　　金：食品安全监测与预警湖南省重点实验室开放基金课题项目(2020KFJJ13);湖南省市场监督管理局科技计划项目(2020KJJH03);长沙市自然科学基金项目(B2020277);食品安全快速检测与溯源关键技术研究与示范项目(2020SK2128)。

摘　　要：为快速、准确地检测出乳制品中赭曲霉毒素A(ochratoxin A,OTA)产生菌,以便评估乳制品中OTA污染的潜在风险,根据已阐明的OTA生物合成途径,选择与OTA产生有关的卤化酶基因区域设计兼并引物,建立针对OTA产生菌的聚合酶链式反应(polymerase chain reaction,PCR)检测方法。对1株产OTA(赭曲霉菌(Aspergillus ochraceus))和5株不产OTA的真菌(冠突散囊菌(Aspergillus cristatus)、黄曲霉菌(Aspergillus flavus)、串珠镰孢菌(Fusarium moniliforme)、土壤伊莎酵母(Issatchenkia terricola)、酿酒酵母(Saccharomyces cerevisiae))DNA进行PCR检测验证。结果表明:产OTA的真菌DNA能扩增出相应的条带,而不产OTA的真菌DNA则无法扩增出相应的条带,本方法在污染的乳制品中也具有较高的准确性和灵敏度。To quickly and accurately evaluate the risk of ochratoxin A(OTA)pollution in dairy products,a method to detect fungi capable of producing OTA in dairy products was developed using gene probe scanning technology.According to the elucidated biosynthesis pathway of OTA,the halogenase gene related to OTA production was selected to design degenerate primers to establish a polymerase chain reaction(PCR)method for the detection of OTA-producing fungi.One OTAproducing strain(Aspergillus ochraceus)and five strains not able to produce OTA(Aspergillus cristatus,Aspergillus flavus,Fusarium moniliforme,Issatchenkia terricola,and Saccharomyces cerevisiae)were selected to test the PCR method.It was found that DNA from the OTA-producing strain but not the non-OTA producers could be amplified by PCR.Moreover,this method had high accuracy and sensitivity for detecting polluted dairy products.

关 键 词：基因探针 高通量 产毒真菌 赭曲霉毒素A 乳制品 风险评估 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]