宏基因组测序技术检测胎菊饮片表面微生物污染方法研究

目的 建立基于宏基因组测序技术检测胎菊表面微生物含量的方法，检测市售中药饮片胎菊微生物污染情况，探索微生物污染途径和来源。方法 利用宏基因组测序比较市售胎菊样品培养组与非培养组之间的表面微生物检出差异，并运用宏基因组测序技术检测2款胎菊商品的表面微生物含量。检测杭州原种场和桐乡新鲜胎菊样品的表面微生物种类，并比较新鲜样品与商品表面微生物种类及相对含量差异。结果 相较于培养组，宏基因组测序技术可以从非培养组中检出更多真菌和古菌。2款胎菊商品表面均检出条件致病菌，饮片表面的微生物与同产地新鲜胎菊样品表面微生物组成相近，表明污染可能来自于种植环境。结论 市售胎菊商品中存在微生物污染情况，宏基因组测序方法可以有效检测胎菊饮片表面微生物，可作为中药饮片微生物污染监测、中药饮片污染微生物数据库构建的有力工具，进而为提高中药饮片的用药安全奠定技术基础。

Study of Metagenomic Sequencing Technology to Detect Microbial Contamination on the Surface of Chrysanthemi Flos

OBJECTIVE To establish a method based on metagenomic sequencing technology to detect the microbial content on the surface of Chrysanthemi Flos, detect the microbial contamination of Chrysanthemi Flos in commercially available Chinese medicine decoction pieces, and explore the ways and sources of microbial contamination. METHODS Metagenomic sequencing was used to compare the surface microbial detection differences between the cultured group and the non-cultured group of commercially available Chrysanthemi Flos samples, and the metagenomic sequencing technology was used to detect the surface microbial content of the two Chrysanthemi Flos products. Detect the surface microbe species of fresh Chrysanthemi Flos samples from Hangzhou original seed farm and Tongxiang county, and compare the species and relative content of microbes on the surface of fresh samples and commercial products. RESULTS Compared with cultured group, more fungi and archaea could be detected in the non-cultured group by metagenomic sequencing technology. Conditional pathogens were detected on the surface of both two commercial brands. Compared with fresh samples from different producing areas, more similar microbial community structure was found between the commercial samples and the fresh ones from the same producing area, which suggested that the microbial contamination mainly came from their original place. CONCLUSION Microbial contamination is present in commercial Chrysanthemi Flos samples, and the use of metagenomic sequencing in cultivation-independent samples can detect the microbial community structure. Thus, metagenomic sequencing can be considered as a powerful tool for the monitoring of microbial contamination and improving the drug safety of the Chinese herbal medicine.