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目的建立食品中致癌真菌污染的培养分离结合实时荧光PCR检测技术,对厦门地区一些可能污染真菌的食品进行监测。方法按照国家标准方法GB/T4789.15-2003进行食品检样的真菌分离培养,并采用涂片染色及镜检方法初步分析真菌种类。在分析比较真菌产毒素相关基因的基础上,依据黄曲霉和寄生曲霉产毒素相关基因的保守区序列设计了两组特异性引物及荧光标记探针,采用液氮研磨结合CTAB法提取真菌DNA,应用实时荧光PCR技术对分离的曲霉菌进行了基因检测。结果从11种食品检样中共分离到35株曲霉菌、2株镰刀菌和6株青霉菌而从糯米鸡、咖喱角和春卷等三种食品中分离的13株曲霉菌含有产毒素相关基因(即为可能的致癌菌株),占全部35株曲霉菌的37.14%。结论提示厦门市食品中致癌真菌污染的比例较高(27.27%),应引起重视。
Objective To establish a culture separation of carcinogenic fungi in foodstuffs and detect real-time fluorescent PCR detection techniques to monitor some foods that may contaminate fungi in Xiamen area. Methods According to the national standard method GB / T4789.15-2003, the fungi isolated from food samples were isolated and cultured. The fungal species were analyzed by smear staining and microscopic examination. Based on the analysis of genes related to fungal toxin production, two sets of specific primers and fluorescently labeled probes were designed according to the conserved region sequences of Aspergillus flavus and Aspergillus parasiticus toxins. The fungal DNA was extracted by liquid nitrogen grinding combined with CTAB, The isolated Aspergillus was tested for gene using real-time PCR. Results Thirty-five Aspergillus, two Fusarium and six Penicillium strains were isolated from 11 food samples and 13 Aspergillus strains isolated from three kinds of foods such as glutinous rice, curry angle and spring roll contained toxin-producing genes That is, possible carcinogenic strains), accounting for 37.14% of all 35 aspergillus. The conclusion suggests that the proportion of carcinogenic fungi in Xiamen food is high (27.27%), which should be paid more attention.