摘要
目的了解不同环境因素对肠杆菌科细菌生物量的影响和耐药基因的表达差异及环境因素对肠杆菌科细菌种群的影响,为探讨环境因素对肠杆菌科细菌的耐药表型及tetA的耐药影响机制提供依据。方法以野生型的肺炎克雷伯细菌为主要研究对象(大肠埃希氏菌为对比),经如下条件进行培养:盐度(0.00%、0.90%、1.80%、2.70%)、紫外处理(于无菌工作台30 W紫外灯处理时间为0、10、20、30 min),然后提取细菌总RNA、经反转录及Realtime-PCR,再根据2^(-ΔΔCt)计算tetA基因相对表达量。结果当盐度为中等浓度时(1.80%)基因tetA的表达量最大,且高盐度及高紫外照射时长(照射30 min)时,目的基因表达量相对较小;其中盐度及紫外培养的组间差异有统计学意义(P<0.01)。结论该海洋来源的多重耐药肺炎克雷伯菌能耐一定盐度,但在高盐度和高紫外照射时长时细菌的tetA基因的表达受到一定的限制。
Objective To understan the influence of different environmental factors on the biomass of enterobacteriaceae, and the different expression of antibiotic-resistant genes, so as to provide a basis for the influence of environmental factors on the drug-resistant phenotype of enterobacteriaceae and the mechanism of tetA. Methods The wild type klebsiella pneumoniae was taken as the research object ( escherichia coli as the comparison). The klebsiella pneumoniae were cultured under different conditions: salinity (0. 00%, 0. 90%, 1. 80%, 2. 70%) and ultraviolet treatment (30 W ultraviolet with the time of 0,10,20,30 min). Then total RNA was extracted,and treating with reverse transcription and Realtimepcr. The method of 2-△△Ct formula was used to calculate relative expression of genes. Results When the salinity was moderate (1. 80%),the expression of gene tetA was the highest,and the expression of tetA was relatively lower when the salinity and the ultraviolet exposure was high (30 min). Among them, the differences among groups in salinity ( P < 0. 01) and ultraviolet ( P <0. 01) were statistically significant. Conclusion The multi-drug resistant klebsiella pneumoniae from this marine source can tolerate certain salinity,but the expression of tetA gene of the bacteria is limited to a certain extent at high salinity and high ultraviolet duration.
作者
韦孟咪
梁枫
胡高垚
周鹤
许士兰
林妍
吴纪贞
龙文芳
WEI Meng-mi;LIANG Feng;HU Gao-yao;ZH()U He;XU Shi-Ian;LIN Yan;WU Ji-zhen;LONG Wen-fang(School of Public Health , Hainan Medical University , Haikou, Hainan ,571199,China)
出处
《预防医学论坛》
2019年第5期328-330,共3页
Preventive Medicine Tribune
基金
国家自然科学基金(81460487),海水浴场肠杆菌科细菌耐药影响机制及抗性基因转移研究
2018年度海南省教育厅研究生创新项目(hys2018266)
国家大学生创新课题(201611810056)
海水浴场肠杆菌的耐药调查及抗性基因的流行研究
海南省大学生创新课题(2017102)海水浴场的从业人群肠杆菌科细菌耐药的影响因素研究
