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基于氨基酸的离子液体对口腔感染粪肠球菌致病基因的分离及其实时 PCR 表达。

Isolation of Enterococcus faecalis pathogenic genes isolated from oral infection by the effect of ionic liquids based on amino acid and its expression by real-time PCR.

机构信息

Department of Microbiology, College of Bioscience, Arak Islamic Azad University, Arak, Iran.

Department of Microbiology, School of Basic Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran.

出版信息

Folia Microbiol (Praha). 2022 Oct;67(5):777-784. doi: 10.1007/s12223-022-00972-7. Epub 2022 May 19.

Abstract

Enterococcus faecalis is an important factor in nosocomial infections. The aim of this study was the isolation of the pathogenic genes of Enterococcus faecalis in mouth infection and the study of the expression of these genes by real-time PCR. In this study, 60 isolates of E. faecalis were isolated from oral infections. The presence and frequency of cyl, hyl, and esp genes and their expression by ionic liquid were evaluated using real-time PCR. MIC was determined by broth dilution method and biofilm production was measured, then biofilm inhibition ability and cytotoxicity test were performed by MTT method. The esp, cyl, and hyl genes were observed in 10, 11, and 2 isolates, respectively. cyl gene with the highest frequency of expression in the treated group was reduced by 1.14% under the influence of ionic liquid with methionine base. The results of MIC and Sub MIC concentrations were obtained with the effect of ionic liquid including 125 and 225 µg/mL, respectively. Amino-acid-based ionic liquids can also reduce biofilm production at sub-MIC concentrations (P < 0.05), and changes in cytotoxicity at different concentrations and over time are significant (P-value < 0.001). E. faecalis strains are genetically diverse and this indicates the polyclonal prevalence of strains in clinical specimens. Combination treatment of ionic liquid with common antimicrobial drugs has good antibacterial effects against Enterococcus species, and ionic liquid with a minimum dose can be a good alternative to single-drug treatment of Enterococcus infections.

摘要

粪肠球菌是医院感染的重要因素。本研究的目的是分离口腔感染粪肠球菌的致病基因,并通过实时 PCR 研究这些基因的表达。本研究从口腔感染中分离出 60 株粪肠球菌。采用实时 PCR 法检测 cyl、hyl 和 esp 基因的存在和频率及其在离子液体中的表达。采用肉汤稀释法测定 MIC,测定生物膜生成量,采用 MTT 法进行生物膜抑制能力和细胞毒性试验。观察到 10、11 和 2 株分离物中分别存在 esp、cyl 和 hyl 基因。在离子液体甲硫氨酸的影响下,处理组 cyl 基因的表达频率降低了 1.14%。MIC 和亚 MIC 浓度的结果分别为离子液体的 125 和 225μg/ml 所获得。含氨基酸的离子液体也可以降低亚 MIC 浓度下的生物膜生成量(P<0.05),不同浓度和时间点的细胞毒性变化有显著差异(P 值<0.001)。粪肠球菌菌株具有遗传多样性,这表明临床标本中菌株的多克隆流行。离子液体与常用抗菌药物联合治疗对肠球菌属具有良好的抗菌作用,最小剂量的离子液体可以作为肠球菌感染单一药物治疗的良好替代品。

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