Application of amplicon sequencing technology to identify contaminating microbes in Chinese patent drugs
Objective:To evaluate the application value of amplicon sequencing in identification of drug-contami-nating microbes in Chinese patent drugs.Methods:Both culturing method and Amplicon Sequencing technology were used to identify bacteria and fungi in 16 batches of Chinese patent drugs.The microbial colonies isolated by culturing medium were identified by 16srDNA/ITS sequencing and MALID-TOF-MS.The amplicon sequencing reads were clustered to OTU(Operational Taxonomic Units)by 97％similarity and BLAST in NCBI to get species taxonomy.Alpha and beta diversity index were used to analyze the microbial community composition in and between batches of drugs.Results:Fifteen species of bacteria and 3 species of fungi were isolated by culturing method,while 75 species of bacteria and 78 species of fungi were identified by amplicon sequencing.Between pre-cultured and non-cultured drug groups,the α diversity index and abundance of most dominant microbial genus were not changed significantly(P＞0.05).Beta diversity analysis showed that microbial diversity between different drugs are more than the same drugs between different pre-treated groups.Conclusion:This study revealed that amplicon sequencing method can provide more comprehensive information on the microbial community composition than tradi-tional culture method.It contributes great value to quick inspection and monitor for microbial contamination in chines patent drugs.
amplicon sequencingChinese patent drugsmicrobial contaminationOTU clustering,alpha and beta diversity
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