基于ITS序列的化妆品中污染真菌检测与鉴定

Application of ITS sequences in detection and identification of Fungi in cosmetics

目的对化妆品中污染的微生物进行分子生物学方法鉴定,明确污染化妆品的真菌种类。方法对化妆品中的污染真菌进行分离培养,通过扩增ITS序列并测序,同时观察菌落形态及在光学显微镜下菌株形态,对其进行鉴定。结果从微生物污染化妆品样品中分离培养到五株污染真菌,观察其在马铃薯葡萄糖琼脂培养基上的生长形态和在光学显微镜下观察其菌落形态,并通过扩增ITS序列对其进行鉴定,结果化妆品所污染的真菌分别为极细链格孢霉、短帚霉、青霉和枝状枝孢霉。结论本研究通过利用分子生物学手段对这些菌株进行了鉴定,明确了化妆品中易污染真菌的种类,为弄清化妆品真菌污染源,做好真菌污染的控制和防范工作提供了依据,同时为致病真菌检测相关规范的建立奠定了基础。

Objective Using the molecular biology methods to classify and identify fungi in cosmetics. Methods The fungi in cosmetics were isolated and cultured to observe the bacteria morphology by amplification of ITS sequence. Results Five fungi strains were isolated from microbe - contaminated cosmetics samples. Then the 5 isolates were inoculated onto Potato Dextrose Agar （PDA） culture medium to observe the growth morphology and colony morphology under optical microscope. With amplification of ITS sequence, the isolates were identified as Alternaria tenmissima, Scopulariopsis brevicaulis, Penicillium chrysogenum and Cladosporium cladosporioides. Conclusion In this study, molecular biological methods were used for the identification of fungi species in polluted cosmetics, which can contribute to clarify fungi pollution sources of cosmetics and provide the basis for control and prevention of fungi pollution. At the same time, it has also laid the foundation for establishment of the relevant specifications for pathogenic fungus test in cosmetics.