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.
作者
李曲文
李盈盈
郑恩惠
杨劲松
徐海滨
翁顺太
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)
出处
《中国病原生物学杂志》
CSCD
北大核心
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)。
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
Legionella
molecular typing
the analysis of multiple point number variable series repeat sequence
matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS)
