Effect of sub-MIC concentrations of metronidazole, vancomycin, clindamycin and linezolid on toxin gene transcription and production in Clostridium difficile
Effect of sub-MIC concentrations of metronidazole, vancomycin, clindamycin and linezolid on toxin gene transcription and production in Clostridium difficile
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DOI：
10.1099/jmm.0.47739-0
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发表时间：
2008-06-01
影响因子：
3
通讯作者：
Ackermann, Grit
中科院分区：
文献类型：
--
作者：
Gerber, Michael;Walch, Christiane;Ackermann, Grit
艰难梭菌是医院获得性感染性腹泻的主要原因。已知几种抗微生物剂诱导和促进C.艰难梭菌相关性腹泻（CDAD）。研究了甲硝唑（MTR）、万古霉素（货车）、克林霉素（CLI）和利奈唑胺（LZD）对大肠杆菌生长、毒素基因转录和毒素产生的影响。difficile进行了研究。四个C在有和没有亚MIC浓度的MTR、货车、CLI和LZD（0.5x MIC）的情况下培养艰难梭菌菌株，并通过菌落计数测量生长。使用ELISA（对于毒素A）和细胞毒性测定（对于毒素B）检测培养物上清液和超声处理的细胞中的毒素产生。实时PCR用于测量毒素A和B基因的转录。本工作的目的是将联合收割机通过ELISA或细胞培养测定与转录组学分析相结合来分析毒素A和B的产生。在没有抗生素的情况下，四种菌株显示出相似的生长和不同水平的毒素产生。无菌对照组在细菌生长的后期达到平台期时显示毒素产生，而暴露于无菌的菌株显示毒素产生较早。除CLI外，所有使用的抗生素都增加了毒素基因的转录速率。本研究结果表明，亚MIC浓度的抗生素可引起梭菌主要毒力因子基因转录的改变。很难本研究建立了一种新的毒素基因转录组学分析方法。很难
Clostridium difficile is the major cause of hospital-acquired infectious diarrhoea. Several antimicrobials are known to induce and promote C. difficile-associated diarrhoea (CDAD). The impact of metronidazole (MTR), vancomycin (VAN), clindamycin (CLI) and linezolid (LZD) on growth, toxin gene transcription and toxin production in C. difficile was investigated. Four C. difficile strains were grown with and without sub-MIC concentrations of MTR, VAN, CLI and LZD (0.5x MIC) and growth was measured by colony counts. Toxin production was detected using ELISA (for toxin A) and a cytotoxicity assay (for toxin B) in culture supernatants and also in sonicated cells. Real-time PCR was used to measure transcription of the toxin A and B genes. The aim of this work was to combine analysis of toxin A and B production by ELISA or cell culture assay with transcriptomic analysis. The four strains showed similar growth and different levels of toxin production in the absence of antibiotics. An antibiotic-free control showed toxin production at a late stage when the plateau phase of bacterial growth was reached, whereas antibiotic-exposed strains showed earlier toxin production. All of the antibiotics used except CLI increased the transcription rate of toxin genes. The findings of this study show that sub-MIC concentrations of antibiotics can cause changes in gene transcription of the major virulence factors of C. difficile. This study describes a new method for transcriptomic analysis of toxin genes in C. difficile.