首页> 中文期刊> 《吉林大学学报(医学版)》 >亚抑菌浓度亚胺培南对MRSA体外增殖及毒力相关基因mRNA表达水平的影响

亚抑菌浓度亚胺培南对MRSA体外增殖及毒力相关基因mRNA表达水平的影响

         

摘要

Objective:To explore the effect of sub-inhibitory concentration of imipenem on the bio-activities of methicillin resistant Staphylococcus aureus(MRSA) and illuminate the inhibitory effects of carbapenem antibioty on the activity of MRSA and their mechanisms,and to provide the basis for using the carbapenem antibiotics in the treatment of MRSA infection.Methods:Five ST239 type of MRSA clinical isolates were selected and were co-cultured with 1/10 and 1/2 minimal inhibitory concentration (MIC) of imipenem for 1.5,6.0 and 12.0 h,which were divided into control group(no imipenem),1/10MIC group(1/10MIC of imipenem),and 1/2MIC group(1/2MIC of imipenem).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes fibronectin A(fnbA),staphylococcal protein A(spa),α-hemolysin(Hla),leukocidin D(lek-D),leukocidin E(lek-E),and enterotoxin A in various groups;Spectrophotometry was used to detect the proliferation activity of MRSA strains in various groups in vitro.Results:After co-culture for 6.0 and 12.0 h,the proliferation activities of 5 trains of MRSA in 1/2MIC group in vitro were significantly decreased compared with control group (P<0.01).The relative mRNA expression levels of 6 virulence-related genes of 5 strains of MRSA in 1/10MIC and 1/2MIC groups were significantly decreased compared with control group(P<0.01).After co-culture for 12.0 h,the mRNA expressions of all the tested virulence-related genes were not found.Conclusion:The sub-inhibitory concentration of imipenem shows obviously inhibitory effect on the mRNA expressions of multiple virulence-related genes of ST 239 type of MRSA strains,and higher concentration of imipenem can suppress the proliferation of MRSA strains in vitro.Imipenem shows the potential value in the treatment of the severe MRSA infected patients.%目的:探讨亚抑菌浓度亚胺培南对耐甲氧西林金黄色葡萄球菌(MRSA)生物学活性的影响,阐明碳青霉烯类抗生素对MRSA活性的抑制作用及其机制,为临床应用碳青霉烯类抗生素治疗MRSA感染提供依据.方法:选取5株ST239型MRSA临床分离株,采用1/10和1/2最小抑菌浓度(MIC)亚胺培南与其体外共培养1.5、6.0和12.0 h,分为对照组(培养液中不加亚胺培南)、1/10MIC组(培养液中添加1/10MIC浓度亚胺培南)和1/2MIC组(培养液中添加1/2MIC浓度亚胺培南).采用荧光定量PCR方法检测各组MRSA株毒力相关基因纤维黏连蛋白A(fnbA)、葡萄球菌蛋白A(spa)、α溶血素(hla)、白细胞毒素D(lek-D)和E(lek-E)、肠毒素A(sea)mRNA相对表达水平,分光光度法检测各组MRSA株体外增殖活性.结果:与对照组比较,体外共培养6.0和12.0 h时,1/2MIC组MRSA株增殖活性明显降低(P<0.01);培养1.5和6.0 h时,6个MRSA毒力相关基因mRNA相对表达水平均明显低于对照组(P<0.01);培养12 h时,1/10MIC和1/2MIC浓度组MRSA株各毒力相关基因mRNA表达未能检测到.结论:亚抑菌浓度亚胺培南对ST239型MRSA多种毒力相关基因mRNA表达具有明显抑制效应,高浓度亚胺培南可抑制MRSA的体外增殖,提示亚胺培南对于重症MRSA感染患者具有潜在的应用价值.

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