基于MALDI-TOF-MS和MLVA技术对军团菌的鉴定和分型研究  

Identification and typing of Legionella based on MALDI-TOF-MS and MLVA techniques

作　　者：李曲文[1,2,3] 李盈盈 郑恩惠 杨劲松[1] 徐海滨[1] 翁顺太[1] LI Qu-wen;LI Ying-ying;ZHENG En-hui;YANG Jin-song;XU Hai-bin;WENG Shun-tai(Fujian Center for Disease Control and Prevention(Fujian Provincial Key Laboratory of Zoonosis Research),Fuzhou 350001,China;Professional Teaching and Practice Base,School of Public Health,Fujian Medical University,Fuzhou 350001,China;Department of Laboratory Medicine,Fujian Medical University,Fuzhou 350001,China;School of Public Health,Fujian Medical University,Fuzhou 350001,China)

机构地区：[1]福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,福建福州350001 [2]福建医科大学公共卫生学院专业教学实习基地 [3]福建医科大学检验系 [4]福建医科大学公共卫生学院

出　　处：《中国病原生物学杂志》2021年第10期1139-1142,共4页Journal of Pathogen Biology

基　　金：福建省自然科学基金项目(No.2019J01093);福建省预防医学研究院建设项目(No.2019Y2001);福建省传染病病原谱监测研究项目(No.2017ZX10103008004)。

摘　　要：目的了解福建省4个地区军团菌的基因特征与分布情况。方法采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)对2013-2020年分离自福建省4个城市的32株9个血清型(Lp1、Lp3、Lp4、Lp5、Lp6、Lp7、Lp8、Lp9、L. micdadei)军团菌进行鉴定,结合多位点串联重复序列分析(MLVA)检测分析其中8个位点(lpsm1,lpsm3,lpsm13,lpsm17,lpsm19,lpsm33,lpsm34,lpsm35)对其中的13株Lp1进行分型。结果 32株军团菌均可通过MALDI-TOF MS检测到属种水平,鉴定分值平均值为2.04±0.16;通过蛋白聚类可以分为A、B、C 3个群;对其中的13株Lp1菌株进行MLVA分型,菌株间相似度为40.9%~87.5%,按照100%的相似水平可分为13个MLVA型,无优势MLVA型。Lp1 2013-25(7号菌)和Lp1 2013-14(9号菌)在MALDI-TOF MS和MLVA两种聚类均显示亲缘关系较近,并与其他菌株均存在差异。结论 MALDI-TOF MS技术可应用于军团菌的快速鉴定,但MALDI-TOF MS与MLVA用于军团菌的分子分型并不完全一致,可联合应用分析军团菌的群体遗传关系。Objective To investigate the genetic characteristics and distribution of Legionella in four different regions of Fujian Province. Methods Matrix-assisted laser analytical ionization time of flight mass spectrometry(MALDI-TOF MS) was used to identify and detect 32 strains of 9 serotypes(Lp1, Lp3, Lp4, Lp5, Lp6, Lp7, Lp8, Lp9, and L. micdadei) of Legionella isolates from four cities in Fujian Province from 2013 to 2020. Based on simple and rapid detection with MALDI-TOF MS, the direct detection of Legionella and Legionella bacterial protein was compared and analyzed. Multiple locus variable-number tandem repeats analysis(MLVA) was also performed, and 13 strains of Lp1 were genotyped at 8 loci(lpsm1, lpsm3, lpsm13, lpsm17, lpsm19, lpsm33, lpsm34, and lpsm35). Results Thirty-two strains of Legionella were identified at the genus and species level using MALDI-TOF MS. The average identification score was 2.04 ± 0.16. The strains were divided into three groups-A, B, and C-based on protein clustering. After MLVA typing, the similarity of 13 LP1 strains ranged from 40.9% to 87.5%. Based on a similarity of 100%, 13 MLVA types were classified, and no dominant MLVA types were found. Lp1 2013-25(strain 7) and Lp1 2013-14(strain 9) were closely related in MALDI-TOF MS and MLVA clusters and differed from other strains. Conclusion MALDI-TOF MS can be used to rapidly identify Legionella. MALDI-TOF MS and MLVA results are not completely consistent when used to molecularly type Legionella, so they can be combined to analyze the genetic relationship among Legionella strains.

关 键 词：军团菌 分子分型 多位点数目可变串联重复序列分析 基质辅助激光解吸电离飞行时间质谱 

分 类 号：R378[医药卫生—病原生物学]