15种中药材表面污染真菌的分离与分子鉴定

Separation and molecular identification of fungal contamination on surface of 15 Chinese herbal medicines

为了了解中药材受真菌污染的状况,探索中药材表面污染真菌的快速鉴定方法。利用PDA培养基进行中药材污染真菌的分离与纯化。根据真菌基因组ITS序列设计特异引物,通过PCR扩增和序列分析进行菌株鉴定。从15种湖北市售中药材上分离得到50株真菌,污染率为93.3%。利用分子鉴定方法成功鉴定了其中的27株。中药材受真菌污染现象较为普遍,各菌属污染中药材情况不同,其中以曲霉属真菌的污染较严重,存在潜在的风险。该实验所设计的引物通用性较好,可用于中药材表面污染真菌的鉴定。

Objective： To evaluate fungal contamination on the surface of Chinese herbal medicines and explore an appropriate method for fast and efficient identification of contaminant fungi. Method： Chinese herbal medicines were first washed and the washing solution was plated onto potato dextrose agar （PDA） to obtain the pure isolates. For molecular identification, two new pairs of specific primers were designed according to ITS region of fungi genome sequences. The strains were identified through polymerase chain reaction （PCR） and sequence analysis. Result： Fifty fungal strains were obtained from the surface of 15 Chinese herbal medicines with the per- cent of contaminated samples of 93.3%. Twenty-seven strains among them were successfully identified. Conclusion： Fungal contami- nation on the surface of Chinese herbal medicines is quite common. Although different fungal species were isolated, the genus Aspergil- lus was the predominant. The primer pairs developed in this study are compatible and can be used to identify fungal species from the surface of Chinese herbal medicines.