双J管生物被膜细菌耐药及其相关基因研究
发布时间：2018-10-22 19:50
【摘要】：1.双J管感染生物被膜细菌临床分布及耐药特性的研究 目的 筛选双J管感染生物被膜菌,了解其临床分布,通过对其生物被膜菌与浮游菌耐药差异性的分析,探讨细菌生物被膜对抗生素的耐药特性,尝试了解生物被膜菌在体内较“真实”的耐药性。 方法 (1)双J管感染生物被膜菌的筛选鉴定及临床分布：收集湘雅三医院2009年2月～2009年7月92例患者双J管。筛选生物被膜菌株和相应浮游株；了解尿路感染生物被膜细菌的临床分布。 (2)临床耐药差异性分析：浮游菌与双J管生物被膜菌在MH培养基上耐药性分析以及生物被膜阳性菌在泊洛沙姆(Poloxamer,F-127)培养基和普通MH培养基上耐药差异性分析。 结果 (1)我院92例患者双J管中经刚果红试验筛选出生物被膜阳性菌41株,阳性率为45%,经鉴定,以肠球菌属(46.3%)和革兰阳性的葡萄球菌属(38.8%)占多数。葡萄球菌属中又以血浆凝固酶阴性菌为主,如表皮葡萄球菌、溶血葡萄球菌和人型葡萄球菌；粪肠球菌位居整个感染菌的第一位。而革兰阴性菌只占生物被膜菌的14.6%,包括铜绿假单胞菌和大肠杆菌。 (2)生物被膜菌组与相应的浮游菌组在普通MH培养基上药敏率的比较差异无统计学意义(P0.05)；生物被膜菌在Poloxamer培养基和MH培养基上,大多数药物的耐药率差异有统计学意义(P0.05),且生物被膜菌在Poloxamer培养基中耐药率较MH培养基更高。 结论 本院双J管感染生物被膜菌在临床上以葡萄球菌属和肠球菌属为主；生物被膜菌与浮游菌在体外耐药性分析未见明显差异,Poloxamer培养基有可能模拟出生物被膜菌的生存环境,而且对药物的耐受性更强。 2.粪肠球菌生物被膜形成与其相关基因三者相关性研究 目的 以粪肠球菌为研究对象,探讨粪肠球菌相关基因(明胶酶编码基因gelE、菌毛操纵子ebpA),与肠球菌生物被膜形成的相关性；对抑制生物被膜的形成提供理论依据。 方法 采用逆转录PCR、实时荧光定量PCR方法,对生物被膜和浮游菌组细菌ebpA、gelE两种与生物被膜形成相关的基因其表达进行检测。 结果 ebpA、gelE基因对粪肠球生物被膜菌形成有一定关系。生物被膜菌组ePbpA表达量分别是浮游菌组的2019倍,表明ebPA与促进生物被膜形成有关,生物被膜菌组gelE表达量分别是浮游菌组的1/138,说明gelE是抑制生物被膜形成原因之一 结论 ebpA、gelE基因与粪肠球生物被膜菌形成有一定关系。ebpA与促进生物被膜形成有关,gelE与抑制生物被膜形成有关。
[Abstract]:1. Study on the clinical distribution and drug resistance of biofilm bacteria infected with double J tube objective to screen biofilm bacteria infected with double J tube and understand their clinical distribution. By analyzing the difference of drug resistance between biofilm bacteria and phytoplankton bacteria, the characteristics of bacterial biofilm resistance to antibiotics were discussed, and the "real" drug resistance of biofilm bacteria in vivo was studied. Methods (1) screening, identification and clinical distribution of biofilm bacteria infected with double J tube: 92 patients were collected from February 2009 to July 2009 in Xiangya third Hospital. Screening biofilm strains and corresponding planktonic strains; To understand the clinical distribution of biofilm bacteria in urinary tract infection. (2) difference analysis of clinical drug resistance: resistance analysis of plankton bacteria and biofilm bacteria in MH medium and biofilm positive bacteria in Poloxamer Difference analysis of drug resistance between (Poloxamer,F-127) medium and MH medium. Results (1) 41 strains of biofilm positive bacteria were screened by Congo red test in 92 cases of double J tube in our hospital. The positive rate of biofilm was 45.5%. Most of them were Enterococcus (46.3%) and Gram-positive Staphylococcus (38.8%). In Staphylococcus, plasma coagulase-negative bacteria were the main bacteria, such as Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus hominis, and Enterococcus faecalis was the first infection bacteria. Gram-negative bacteria only accounted for 14.6of biofilm bacteria, including Pseudomonas aeruginosa and Escherichia coli. (2) there was no significant difference in drug sensitivity between biofilm bacteria group and corresponding phytoplankton group on MH medium (P0.05). The drug resistance rate of biofilm bacteria in Poloxamer medium and MH medium was significantly different (P0.05), and the drug resistance rate of biofilm bacteria in Poloxamer medium was higher than that in MH medium. Conclusion the biofilm bacteria infected with double J tube in our hospital are mainly Staphylococcus and Enterococcus. There was no significant difference between biofilm bacteria and phytoplankton bacteria in drug resistance analysis in vitro. The Poloxamer medium might simulate the living environment of biofilm bacteria, and its tolerance to drugs was stronger. 2. Study on the relationship between biofilm formation and related genes of Enterococcus faecalis objective to study Enterococcus faecalis. To investigate the correlation between Enterococcus faecalis related genes (gelatinase encoding gene gelE, pili operon ebpA), and the formation of biofilm of Enterococcus faecalis) and to provide theoretical basis for inhibiting the formation of biofilm. Methods reverse transcription PCR, real-time fluorescence quantitative PCR was used to detect the expression of biofilm and phytoplankton bacterial ebpA,gelE related to biofilm formation. Results ebpA,gelE gene was related to the formation of membranous bacteria in pellets of feces. The expression of ePbpA in the biofilm bacteria group was 2019 times higher than that in the zooplankton group, indicating that ebPA was related to the promotion of biofilm formation. The expression of gelE in the biofilm bacteria group was 1 / 138 of that in the zooplankton group, which indicated that gelE was one of the reasons for inhibiting the formation of biofilm. Conclusion there is a relationship between the ebpA,gelE gene and the formation of biofilm bacteria in fecal pellets. EbpA is associated with promoting the formation of biofilm. The formation of biofilm is related to the formation of biofilm and gelE is related to the inhibition of biofilm formation.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R378
本文编号：2288142
[Abstract]:1. Study on the clinical distribution and drug resistance of biofilm bacteria infected with double J tube objective to screen biofilm bacteria infected with double J tube and understand their clinical distribution. By analyzing the difference of drug resistance between biofilm bacteria and phytoplankton bacteria, the characteristics of bacterial biofilm resistance to antibiotics were discussed, and the "real" drug resistance of biofilm bacteria in vivo was studied. Methods (1) screening, identification and clinical distribution of biofilm bacteria infected with double J tube: 92 patients were collected from February 2009 to July 2009 in Xiangya third Hospital. Screening biofilm strains and corresponding planktonic strains; To understand the clinical distribution of biofilm bacteria in urinary tract infection. (2) difference analysis of clinical drug resistance: resistance analysis of plankton bacteria and biofilm bacteria in MH medium and biofilm positive bacteria in Poloxamer Difference analysis of drug resistance between (Poloxamer,F-127) medium and MH medium. Results (1) 41 strains of biofilm positive bacteria were screened by Congo red test in 92 cases of double J tube in our hospital. The positive rate of biofilm was 45.5%. Most of them were Enterococcus (46.3%) and Gram-positive Staphylococcus (38.8%). In Staphylococcus, plasma coagulase-negative bacteria were the main bacteria, such as Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus hominis, and Enterococcus faecalis was the first infection bacteria. Gram-negative bacteria only accounted for 14.6of biofilm bacteria, including Pseudomonas aeruginosa and Escherichia coli. (2) there was no significant difference in drug sensitivity between biofilm bacteria group and corresponding phytoplankton group on MH medium (P0.05). The drug resistance rate of biofilm bacteria in Poloxamer medium and MH medium was significantly different (P0.05), and the drug resistance rate of biofilm bacteria in Poloxamer medium was higher than that in MH medium. Conclusion the biofilm bacteria infected with double J tube in our hospital are mainly Staphylococcus and Enterococcus. There was no significant difference between biofilm bacteria and phytoplankton bacteria in drug resistance analysis in vitro. The Poloxamer medium might simulate the living environment of biofilm bacteria, and its tolerance to drugs was stronger. 2. Study on the relationship between biofilm formation and related genes of Enterococcus faecalis objective to study Enterococcus faecalis. To investigate the correlation between Enterococcus faecalis related genes (gelatinase encoding gene gelE, pili operon ebpA), and the formation of biofilm of Enterococcus faecalis) and to provide theoretical basis for inhibiting the formation of biofilm. Methods reverse transcription PCR, real-time fluorescence quantitative PCR was used to detect the expression of biofilm and phytoplankton bacterial ebpA,gelE related to biofilm formation. Results ebpA,gelE gene was related to the formation of membranous bacteria in pellets of feces. The expression of ePbpA in the biofilm bacteria group was 2019 times higher than that in the zooplankton group, indicating that ebPA was related to the promotion of biofilm formation. The expression of gelE in the biofilm bacteria group was 1 / 138 of that in the zooplankton group, which indicated that gelE was one of the reasons for inhibiting the formation of biofilm. Conclusion there is a relationship between the ebpA,gelE gene and the formation of biofilm bacteria in fecal pellets. EbpA is associated with promoting the formation of biofilm. The formation of biofilm is related to the formation of biofilm and gelE is related to the inhibition of biofilm formation.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R378
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