检验医学与临床2024，Vol.21Issue(9)：1224-1228,1234,6.DOI:10.3969/j.issn.1672-9455.2024.09.008
3种方法对1株临床疑难菌的鉴定及结果比较
Identification of a clinically difficult bacterial strain by three methods and comparison of results
摘要
Abstract
Objective MALDI-TOF MS,16 S rRNA sequence analysis,and whole genome sequencing(WGS)were applied to identify a strain of suspected Halomonas isolated from a clinical specimen and to com-pare the results of the three methods.Methods MALDI-TOF MS was used to collect protein profiles of the bacteria to be tested,and the identification of microorganisms was achieved by comparing the characteristic peaks of the profiles.The 16 S rRNA gene sequencing was performed on the bacteria to be tested,and the se-quencing results were compared with database.Based on the genomic sequences obtained from WGS,a com-parative analysis was performed,including the calculation of Average Nucleotide Identity(ANI)by ANIm and ANIb,and the comparison with the Genome Taxonomy Database(GTDB).Phylogenetic trees were construc-ted based on 16 S rRNA sequences and housekeeping gene sequences from WGS,as well as cluster analysis based on COG and KEGG functions.Results MALDI-TOF MS resulted in the identification of the strain to be tested as Halomonas hamiltonii.The 16 S rRNA gene sequence comparison results showed that the similari-ty between the bacteria to be tested and the three Halomonas species(Halomonas hamiltonii,Halomonas stevensii and Halomonas johnsoniae)were all above 99%,and the difference in their similarity was so small that accurate species identification could not be performed.The gene sequencing comparison based on WGS,phylogenetic tree and cluster analysis showed that the bacterium was more closely related to Halomonas stevensii.Conclusion The results of the three methods on the strain identification of the isolate are different.MALDI-TOF MS is more convenient and fast for species identification,which is very suitable for clinical tes-ting;WGS-based bacterial identification is particularly useful for distinguishing genetically very similar spe-cies.关键词
细菌鉴定/基质辅助激光解吸电离飞行时间质谱/16S rRNA序列/全基因组测序Key words
bacterial identification/MALDI-TOF MS/16 S rRNA sequence/whole-genome sequen-cing分类
医药卫生引用本文复制引用
许一丹,谢成彬,苏喆,王频佳..3种方法对1株临床疑难菌的鉴定及结果比较[J].检验医学与临床,2024,21(9):1224-1228,1234,6.基金项目
四川省大学生创新创业训练计划项目(S202013705085) （S202013705085）
成都医学院研究生创新基金项目(YCX2022-03-17) （YCX2022-03-17）
成都医学院检验医学院自然科学基金(JYZK202206) （JYZK202206）
四川省妇幼保健院菁英人才科研启动项目(ZC6.16). （ZC6.16）
