苏州地区耐左氧氟沙星无乳链球菌临床分离株分子及质谱学特征分析  

Molecular and mass spectrometric characteristics of levofloxacin-resistant streptococcus agalactiae clinically isolated in Suzhou area,China

作　　者：张宇蔺 苏宁 陈海鹏 秦国英 陆文香[1] 徐卫东[1] 袁栎[2] 钟桥[1] ZHANG Yulin;SU Ning;CHEN Haipeng;QIN Guoying;LU Wenxiang;XU Weidong;YUAN Li;ZHONG Qiao(Department of Clinical Laboratory Medicine, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou 215002, Jiangsu;Department of Biochemistry and Molecular Biology, School of Basic Medicine, Nanjing Medical University, Nanjing 211166, Jiangsu, China)

机构地区：[1]南京医科大学附属苏州医院医学检验科,江苏苏州215002 [2]南京医科大学生物化学与分子生物学系,南京211166

出　　处：《临床检验杂志》2020年第10期741-745,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(81971995);江苏省自然科学基金(BK20191179);苏州市“科教兴卫”青年基金(KJXW20170125)。

摘　　要：目的回顾分析苏州地区临床分离左氧氟沙星耐药无乳链球菌菌株分子及蛋白质谱学特征。方法收集2018年1月至2019年1月临床连续分离非重复耐左氧氟沙星无乳链球菌58株,用PCR法及Sanger基因测序方法检测gyrA、parC和parE基因突变、细菌血清型以及多位点序列分型;用ClinPro Tools 3.0软件对左氧氟沙星耐药菌株蛋白质谱特征峰进行筛选并验证。结果测序检出耐左氧氟沙星菌株以携带gyrA_Ser81Leu+parC_Ser79Tyr+parE_His225Tyr突变为主,占44.8%(26/58),以血清Ⅲ型和ST19型为主,并伴有血清Ⅰb型及ST23型流行;耐药菌株蛋白质谱峰分析显示6894、3445、5076、4544以及3721 m/z为主要特征峰。结论本地区耐左氧氟沙星无乳链球菌主要为gyrA_Ser81Leu+parC_Ser79Tyr+parE_His225Tyr突变,且以血清Ⅲ型及ST19型为主,具有以6894 m/z为代表的质谱特征峰。Objective To analyze retrospectively the molecular and Mass spectrometer characteristics of levofloxacin-resistant Streptococcus agalactiae isolated clinically in Suzhou area.Methods A total of 58 isolates of non-repetitive levofloxacin-resistant Streptococcus agalactiae were collected from January 2018 to January 2019.PCR and Sanger gene sequencing methods were used to detect the gene mutations of gyrA,parC and parE,as well as the serotype-associated genes and multi-locus sequence typing.ClinPro Tools 3.0 software was used to screen and verify the levofloxacin-resistant specific peaks of mass spectrometry(MS)of proteomics in the Streptococcus agalactiae isolates.Results The results of sequencing showed that levofloxacin-resistant Streptococcus agalactiae strains mainly carried the mutations of gyrA_Ser81Leu+parC_Ser79Tyr+parE_His225Tyr,accounting for 44.8%(26/58).SerotypeⅢand ST19 were the main types of levofloxacin-resistant streptococcus agalactiae isolates accompanied by prevalent serotypeⅠb and ST23.Mass spectrometry profile analysis for streptococcus agalactiae showed that 6894,3445,5076,4544 and 3721 m/z were the dominant specific differential peaks of the levofloxacin-resistant streptococcus agalactiae isolates.Conclusion The gene mutations of levofloxacin-resistant Streptococcus agalactiae isolates in the local area were mainly gyrA_Ser81Leu+parC_Ser79Tyr+parE_His225Tyr with serotypeⅢand ST19,and 6894 m/z was represented as the specific mass spectrometric peak.

关 键 词：无乳链球菌 左氧氟沙星 基质辅助激光解吸电离飞行时间质谱 

分 类 号：R446.5[医药卫生—诊断学]