摘要
目的了解土壤中军团菌的污染状况和分离菌株的分子型别、对抗生素的敏感性和细胞内的生长能力。方法采集北京市公共场所土壤样本123份,采用军团菌GVPC选择性培养基培养法和荧光定量PCR检测法对土壤中军团菌进行检测分析。用小鼠巨噬细胞J774细胞对分离菌株进行菌株致病性研究;用脉冲场凝胶电泳(PFGE)技术对分离菌株进行分型分析;用E-test试纸药敏实验法对分离菌株进行红霉素、阿奇霉素、利福平、左氧氟沙星、莫西沙星5种抗生素敏感性检测。结果荧光定量PCR检测法在花鸟市场和夏季水库样本中检出军团菌,检出阳性率分别为50.00%和24.24%,平板培养法仅在夏季水库样本中检出军团菌,阳性率为6.06%。分离的2株军团菌TR1和TR2均为嗜肺军团菌,血清型分别为5型和8型。2株嗜肺军团菌均具有较高的细胞内生长能力,PFGE分型显示2株菌为不同带型,药敏实验显示2株菌均对红霉素、阿奇霉素、利福平、左氧氟沙星、莫西沙星5种常用抗生素表现敏感性。结论公共场所土壤中可检出嗜肺军团菌,且分离株有较高的细胞内生长能力,提示具有致病性,存在导致军团菌病散发流行或暴发的危险,应加强对公共场所土壤中军团菌的监测。
Objective To understand the contamination of Legionella in soil and the molecular type,sensitivity to antibiotics and intracellular growth ability of isolated strains.Methods A total of 123 soil samples from public places in Beijing were collected.Legionella in soil was detected by GVPC(Legionella selective medium) culture and quantitative real-time PCR detection.The intracellular growth ability of the Legionella isolates were tested by cell culture methods with mouse macrophage J774.The genotypes of the isolates were determined by pulsed-field gel electrophoresis.Antibiotic sensitivity detection of the isolates was conducted with E-test.Results Quantitative PCR assay detected Legionella in flower & bird market samples and reservoir samples in summer.The positive rates were 50.00% and 24.24% respectively.The plate culture method only detected Legionella in reservoir samples in summer.The positive rate was 6.06%.All the two Legionella strains isolated,TR1 and TR2,were L.pneumophila,and the serotypes were type 5 and type 8 respectively.All the two strains of L.pneumophila had high intracellular growth ability.PFGE typing showed two different band types.The drug sensitivity test showed that the two strains were sensitive to erythromycin,azithromycin,rifampicin,levofloxacin and moxifloxacin.TR1 showed resistance to cefuroxime,and TR2 was sensitive to cefuroxime.Conclusions Legionella pneumophila was detected in the soil of public places,and the isolates have higher intracellular growth ability,indicating the pathogenicity of the strains and suggesting the potential of causing legionellosis epidemic.Therefore,we should strengthen the monitoring of Legionella in public places.
作者
张婧
贾雪洋
任红宇
李建国
秦天
Zhang Jing;Jia Xueyang;Ren Hongyu;Li Jianguo;Qin Tian(State Key Laboratory of Communicable Disease Prevention and Control,National Institute for Communicable Disease Prevention and Control,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Modern Research Center for Traditional Chinese Medicine,Taiyuan 030006,Shanxi,China)
出处
《疾病监测》
CAS
2018年第12期1032-1036,共5页
Disease Surveillance
基金
国家自然科学基金(No.81671985)
传染病预防控制国家重点实验室青年创新人才基金(No.2015SKLID508)~~
关键词
土壤
军团菌
脉冲场凝胶电泳
细胞内生长能力
药敏检测
Soil
Legionella
Pulsed-field gel electrophoresis
Intracellular growth ability
Antibiotic sensitivity
