【摘要】：1.雙J管感染生物被膜細(xì)菌臨床分布及耐藥特性的研究 目的 篩選雙J管感染生物被膜菌,了解其臨床分布,通過對(duì)其生物被膜菌與浮游菌耐藥差異性的分析,探討細(xì)菌生物被膜對(duì)抗生素的耐藥特性,嘗試了解生物被膜菌在體內(nèi)較“真實(shí)”的耐藥性。 方法 (1)雙J管感染生物被膜菌的篩選鑒定及臨床分布：收集湘雅三醫(yī)院2009年2月～2009年7月92例患者雙J管。篩選生物被膜菌株和相應(yīng)浮游株；了解尿路感染生物被膜細(xì)菌的臨床分布。 (2)臨床耐藥差異性分析：浮游菌與雙J管生物被膜菌在MH培養(yǎng)基上耐藥性分析以及生物被膜陽性菌在泊洛沙姆(Poloxamer,F-127)培養(yǎng)基和普通MH培養(yǎng)基上耐藥差異性分析。 結(jié)果 (1)我院92例患者雙J管中經(jīng)剛果紅試驗(yàn)篩選出生物被膜陽性菌41株,陽性率為45%,經(jīng)鑒定,以腸球菌屬(46.3%)和革蘭陽性的葡萄球菌屬(38.8%)占多數(shù)。葡萄球菌屬中又以血漿凝固酶陰性菌為主,如表皮葡萄球菌、溶血葡萄球菌和人型葡萄球菌；糞腸球菌位居整個(gè)感染菌的第一位。而革蘭陰性菌只占生物被膜菌的14.6%,包括銅綠假單胞菌和大腸桿菌。 (2)生物被膜菌組與相應(yīng)的浮游菌組在普通MH培養(yǎng)基上藥敏率的比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)；生物被膜菌在Poloxamer培養(yǎng)基和MH培養(yǎng)基上,大多數(shù)藥物的耐藥率差異有統(tǒng)計(jì)學(xué)意義(P0.05),且生物被膜菌在Poloxamer培養(yǎng)基中耐藥率較MH培養(yǎng)基更高。 結(jié)論 本院雙J管感染生物被膜菌在臨床上以葡萄球菌屬和腸球菌屬為主；生物被膜菌與浮游菌在體外耐藥性分析未見明顯差異,Poloxamer培養(yǎng)基有可能模擬出生物被膜菌的生存環(huán)境,而且對(duì)藥物的耐受性更強(qiáng)。 2.糞腸球菌生物被膜形成與其相關(guān)基因三者相關(guān)性研究 目的 以糞腸球菌為研究對(duì)象,探討糞腸球菌相關(guān)基因(明膠酶編碼基因gelE、菌毛操縱子ebpA),與腸球菌生物被膜形成的相關(guān)性；對(duì)抑制生物被膜的形成提供理論依據(jù)。 方法 采用逆轉(zhuǎn)錄PCR、實(shí)時(shí)熒光定量PCR方法,對(duì)生物被膜和浮游菌組細(xì)菌ebpA、gelE兩種與生物被膜形成相關(guān)的基因其表達(dá)進(jìn)行檢測(cè)。 結(jié)果 ebpA、gelE基因?qū)S腸球生物被膜菌形成有一定關(guān)系。生物被膜菌組ePbpA表達(dá)量分別是浮游菌組的2019倍,表明ebPA與促進(jìn)生物被膜形成有關(guān),生物被膜菌組gelE表達(dá)量分別是浮游菌組的1/138,說明gelE是抑制生物被膜形成原因之一 結(jié)論 ebpA、gelE基因與糞腸球生物被膜菌形成有一定關(guān)系。ebpA與促進(jìn)生物被膜形成有關(guān),gelE與抑制生物被膜形成有關(guān)。
[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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R378

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