基于高通量测序的6种甘肃道地药材中药饮片污染微生物群落特征研究  

Community Characteristics of Contaminated Microorganisms in Six Kinds of Gansu Authentic Chinese Herbal Pieces Based on High-Throughput Sequencing

作　　者：孙莺 郭晓鹏[2] 徐勤科 武悦 李莉 李若绮 郭朝晖 SUN Ying;GUO Xiaopeng;XU Qinke;WU Yue;LI Li;LI Ruoqi;GUO Zhaohui(NMPA Key Laboratory for Quality Control of Chinese Materia Medica and Prepared Slices,Gansu Institute for Drug Control,Lanzhou 730000,China;School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China)

机构地区：[1]甘肃省药品检验研究院,国家药监局中药材及饮片质量控制研究重点实验室,兰州730000 [2]兰州理工大学生命科学与工程学院,兰州730050

出　　处：《中国药学杂志》2023年第24期2282-2293,共12页Chinese Pharmaceutical Journal

摘　　要：目的研究6种甘肃道地药材中药饮片中污染微生物的群落特征,建立6种中药饮片负载微生物的种属及其丰度信息数据库,为6种中药饮片微生物限度标准制定提供依据。方法收集市售当归、党参、黄芪、甘草、板蓝根和半夏6种中药饮片各30批,合计180批,参照2020年版《中国药典》,检查180批中药饮片样品的需氧菌总数(total aerobic microbial count,TAMC)、霉菌和酵母菌总数(total combined yeasts andmolds count,TYMC)、耐胆盐革兰阴性菌(bile-tolerant gram-negitive bacteria,BGB)及沙门菌,并基于16S rDNA高通量测序方法研究中药饮片中污染微生物的群落特征。结果180批样品均检出需氧菌、酵母菌和霉菌。lg(TAMC)的均值为3.16,最小值为0.70,最大值为6.30。lg(TYMC)的均值为1.71,最小值为0.70,最大值为3.26。102批检出耐胆盐革兰阴性菌。检出沙门菌2批。通过16S rDNA高通量测序对每种中药饮片进行了微生物多样性分析,建立了6种中药饮片负载微生物的种属及其丰度信息数据库,污染微生物分布于38个门、935个已鉴定属,不同类别中药饮片中的优势菌属具有明显差异。植株自身和加工储存环节引入是饮片微生物群落的两种重要来源。结论研究表明,16S rRNA高通量测序方法比传统培养法能够获得更加全面的中药饮片中污染微生物群落信息,不同类别中药饮片中的优势菌属具有明显差异。中药饮片微生物污染具有一定的致病风险,饮片的微生物限度标准亟待完善,饮片生产加工过程控制亟待加强。OBJECTIVE To study the community characteristics of contaminated microorganisms in six kinds of Gansu authentic Chinese herbal pieces and establish the species and abundance information database of the loaded microorganisms,thus to provide the basis for establishment of the microbial limits for the six kinds of Chinese herbal pieces.METHODS Thirty batches of Chinese herbal pieces of Angelica sinensis(Ang),Codonopsis pilosula(Cod),Astragalus membranaceus(Ast),Glycyrrhiza uralensis(Gly),Isatidis Radix(Isa)and Pinellia ternata(Pin)were collected,180 batches in total.According to the 2020 edition of Chinese Pharmacopoeia,the total number of aerobic bacteria(TAMC),fungi and yeasts(TYMC),bile-tolerant gram-negitive bacteria(BGB)and Salmonella were detected in the samples.The community characteristics of contaminated microorganisms were studied by 16S rDNA high-throughput sequencing method.RESULTS Aerobic bacteria,yeast and mould were detected in all of the 180 batches of samples.The mean value of lg(TAMC)was 3.16,the minimum value was 0.70,and the maximum value was 6.30.The mean value of lg(TYMC)was 1.71,the minimum value was 0.70 and the maximum value was 3.26.Bile-tolerant gram-negitive bacteria were found in 102 batches of samples.Salmonella was detected in two batches.Based on microbial diversity analysis by 16 s rDNA high-throughput sequencing of randomly selected 10 batches of each Chinese herbal pieces,the loaded microbial species in the six kinds of traditional Chinese herbal pieces were identified and their abundance information database was established.It was found that the contaminated microorganism distributed in 38 phylums and 935 known genera,and the dominant bacteria of different categories of Chinese herbal pieces differed significantly.CONCLUSION The research shows that 16S rRNA high-throughput sequencing method can obtain more comprehensive information of contaminated microbial communities in Chinese herbal pieces than traditional culture method.The contaminated microbial communities in different kind

关 键 词：中药饮片 微生物污染 微生物群落结构 微生物群落功能 16S rDNA高通量测序 

分 类 号：R282[医药卫生—中药学] R917[医药卫生—中医学]