摘要
目的对实验室药品微生物限度检查所得真菌进行鉴定溯源。方法采用形态学鉴定、内转录间隔区(ITS序列)分析技术和基质辅助激光解离飞行时间质谱(MALDI-TOF-MS)鉴定法,对从药品中分离纯化的4个菌株和实验环境中采集到的8个菌株进行鉴定和同源性分析,并建立系统发育树。结果ITS测序鉴定结果最大鉴定率均大于99%,结果可靠。其中4株样品检出真菌和1株环境菌的蛋白分子量主要集中在2000~18000 Da范围内,主要质谱峰重叠度高,被鉴定为局限曲霉菌,且亲缘关系非常接近。另4株环境菌和3株环境菌被分别鉴定为聚多曲霉菌和烟曲霉菌。结论样品污染菌来自pH 7.0氯化钠-蛋白胨缓冲液。可使用形态学鉴定、ITS序列分析技术和MALDI-TOF-MS技术对实验室真菌污染进行鉴定和溯源。
Objective To identify and trace the obtained fungi in microbial limit test of laboratory drugs.Methods Identification and homology analysis of four strains of fungi isolated and purified from drugs and eight strains of fungi collected from experimental environment were performed by the morphological identification,internally transcribed spacer(ITS sequence)analysis technology and matrix-assisted laser desorption ionization-time of flight-mass spectrometry(MALDI-TOF-MS)method,and a phylogenetic tree was established.Results The maximum identification rate of ITS sequence was greater than 99%,which was reliable.The protein molecular weights of four strains of fungi from drugs and one strain of fungus from environment were mainly in the range of 2000 to 18000 Da,the overlap degree of their main mass spectra peaks was high,and they were identified as Aspergillus restrictus,with close genetic relationship.Another four and three strains of fungi from environment were identified as Aspergillus sydowii and Aspergillus fumigatus,respectively.Conclusion The fungi in drugs come from sodium chloride-peptone buffer(pH 7.0).We can use morphological identification,ITS sequence analysis technology and MALDI-TOF-MS method to identify and trace the fungal contamination of laboratory drugs.
作者
黄结
李雪玲
朱斌
甘永琦
王涛
零文超
HUANG Jie;LI Xueling;ZHU Bin;GAN Yongqi;WANG Tao;LING Wenchao(Guangxi Institute for Food and Drug Control,Nanning,Guangxi,China 530021)
出处
《中国药业》
CAS
2023年第20期91-94,共4页
China Pharmaceuticals
基金
广西壮族自治区药品监督管理局科研计划项目[桂药监科2021-03(直属)]。
