人面部皮肤来源疑难鉴定细菌的分子鉴定方法探索  

Exploration of molecular identification methods for difficult identification of bacteria originating from human facial skin

作　　者：王丹羽 尚靖[2] 王学硕[1] 骆海朋[1] 李景云[1] 窦越 瞿洪仁 崔生辉[1] 路勇[1] WANG Danyu;SHANG Jing;WANG Xueshuo;LUO Haipeng;LI Jingyun;DOU Yue;QU Hongren;CUI Shenghui;LU Yong(Institute for Cosmetic Control,National Institutes for Food and Drug Control,Beijing 100010,China;School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing,Jiangsu 211100,China)

机构地区：[1]中国食品药品检定研究院化妆品检定所,北京100010 [2]中国药科大学中药学院,江苏南京211100

出　　处：《国际检验医学杂志》2024年第S02期27-32,共6页International Journal of Laboratory Medicine

基　　金：中国食品药品检定研究院中青年发展研究基金(2022C2);中国食品药品检定研究院中青年发展研究基金(2022C7);中国食品药品检定研究院关键技术研究基金(GJJS-2022-8-5)。

摘　　要：目的比较分析全自动生化鉴定系统、MALDI-TOF MS、16S rDNA测序和WGS技术对皮肤来源的24株细菌的鉴定结果,为疑难菌种的准确鉴定提供方法参考。方法针对24株人面部皮肤来源的疑难鉴定菌株,分别采用VITEK 2 Compact生化鉴定、MALDI-TOF MS、16S rDNA系统发育分析以及基于WGS的ANI分析进行菌种鉴定,并评估各鉴定方法的准确度。结果MALDI-TOF MS方法在细菌种水平的鉴定准确度为8.3%,在属水平的鉴定准确度为70.8%;16S rDNA序列分析方法在属水平的鉴定结果与WGS一致,但在种水平的鉴定结果仍存在较大差异。以目的菌株与数据库推荐参考菌种的ANI值为评价标准,仅12株菌(50.0%)能够准确鉴定到种水平。结论针对皮肤来源的疑难鉴定细菌,以VITEK 2 Compact为代表的生化鉴定方法和MALDI-TOF MS的鉴定准确度有限,综合使用16S rDNA测序和WGS等分子鉴定技术能提高鉴定准确度。Objective To compare and analyze the identification results of 24 bacterial strains from skin sources using fully automated biochemical identification system,MALDI-TOF MS,16S rDNA sequencing,and WGS technology,in order to provide methodological references for accurate identification of difficult bacterial strains.Methods For 24 difficult identification strains of human facial skin origin,VITEK 2 Compact bio-chemical identification,MALDI-TOF MS,16S rDNA phylogenetic analysis,and WGS based ANI analysis were used for strain identification,and the accuracy of each identification method was evaluated.Results The MALDI-TOF MS method had an identification accuracy of 8.3%at the bacterial species level and 70.8%at the genus level.The identification results of 16S rDNA sequence analysis method at the genus level were con-sistent with those of WGS,but there were still significant differences in the identification results at the species level.Based on the ANI values of the target strain and the recommended reference strains in the database,only 12 strains(50%)could be accurately identified at the species level.Conclusion For difficult identification of bacteria originating from the skin,biochemical identification methods such as VITEK 2 Compact and MALDI-TOF MS have limited accuracy.The comprehensive use of molecular identification techniques such as 16S rD-NA sequencing and WGS can improve the accuracy of identification.

关 键 词：基质辅助激光解吸电离飞行时间质谱 平均核苷酸一致性 全基因组测序 

分 类 号：R446[医药卫生—诊断学]