本文選題：銅綠假單胞菌 + 主動(dòng)外排系統(tǒng)��； 參考：《福建醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的： 了解本地區(qū)臨床分離銅綠假單胞菌MexAB-OprM型外排系統(tǒng)和膜孔蛋白OprD2的表達(dá)情況及其與銅綠假單胞菌多重耐藥的關(guān)系,為臨床抗感染治療及醫(yī)院感染的監(jiān)測(cè)和控制提供循證醫(yī)學(xué)依據(jù)。 方法： 2008年7月至2009年6月我院住院患者送檢標(biāo)本分離銅綠假單胞菌381株(同一患者同類標(biāo)本分離菌株不重復(fù)計(jì)入)。采用VITEK 2 Compact自動(dòng)微生物分析儀進(jìn)行細(xì)菌鑒定和藥敏測(cè)定。選取多重耐藥(對(duì)三類或以上抗生素耐藥)的銅綠假單胞菌,分別以微量肉湯稀釋試驗(yàn)檢測(cè)MexAB-OprM型主動(dòng)外排系統(tǒng)、熒光定量PCR技術(shù)對(duì)該型外排系統(tǒng)及OprD2進(jìn)行相對(duì)定量測(cè)定,對(duì)高表達(dá)MexAB菌株的調(diào)節(jié)基因mexR進(jìn)行PCR擴(kuò)增,并對(duì)產(chǎn)物進(jìn)行測(cè)序。 結(jié)果： 1.銅綠假單胞菌的感染以兒童和老年人居首,在全院各科室中,ICU患者銅綠假單胞菌分離率最高(14.2%),從感染部位上,主要分布于呼吸道(62.2%)。 2.銅綠假單胞菌的耐藥情況相當(dāng)嚴(yán)峻,對(duì)多數(shù)常用抗生素均有較高的耐藥性,但哌拉西林／他唑巴坦、丁胺卡那霉素等抗生素目前仍是治療銅綠假單胞感染的有效抗菌藥物。 3.質(zhì)子泵抑制劑CCCP對(duì)環(huán)丙沙星的抗菌效果影響較大,有15株銅綠假單胞菌的MIC下降8倍以上,其中6株的CAZ在CCCP存在的情況下MIC也下降8倍以上,另有兩株僅有頭孢他啶的MIC下降8倍以上。亞胺培南的抗菌活性基本不受CCCP的影響。 4.對(duì)頭孢他啶耐藥和敏感的兩組銅綠假單胞菌MexAB-OprM型主動(dòng)外排系統(tǒng)表達(dá)量有統(tǒng)計(jì)學(xué)差異(P0.05),對(duì)環(huán)丙沙星耐藥和敏感的兩組銅綠假單胞菌MexAB-OprM型主動(dòng)外排系統(tǒng)表達(dá)量有統(tǒng)計(jì)學(xué)差異(P0.05),但對(duì)亞胺培南耐藥和敏感的兩組,該型外排系統(tǒng)的表達(dá)量無差別(P0.05)。 5. mRNA相對(duì)定量與頭孢他啶的CCCP抑制試驗(yàn)結(jié)果的一致性較好,環(huán)丙沙星抑制試驗(yàn)陽性菌株中有7株相對(duì)定量不高。 6.膜孔蛋白OprD2基因mRNA的定量檢測(cè)結(jié)果顯示,亞胺培南耐藥組的表達(dá)量低于敏感組,并且兩者差異有統(tǒng)計(jì)學(xué)意義(P0.05) 結(jié)論： 1.銅綠假單胞菌是醫(yī)院感染重要病原菌,顯示多重耐藥,應(yīng)該合理選擇抗生素。 2.銅綠假單胞菌MexAB-OprM型主動(dòng)外排系統(tǒng)的高表達(dá)是其存在多重耐藥的重要機(jī)制。 3. MexAB-OprM型主動(dòng)外排系統(tǒng)的高表達(dá)與銅綠假單胞菌對(duì)CIP和CAZ的耐藥有關(guān),而非IMP的耐藥機(jī)制。 4. OprD2的表達(dá)減少是銅綠假單胞菌對(duì)IMP出現(xiàn)耐藥的重要機(jī)制。
[Abstract]:Objective: To investigate the expression of MexAB-OprM type efflux system and membrane-porin OprD2 in clinical isolates of Pseudomonas aeruginosa and its relationship with multidrug resistance of Pseudomonas aeruginosa, and to provide evidence based medicine basis for clinical anti-infection therapy and monitoring and control of nosocomial infection. Methods: From July 2008 to June 2009 381 strains of Pseudomonas aeruginosa were isolated from inpatients in our hospital. VITEK 2 Compact automatic microbiological analyzer was used to identify bacteria and determine drug sensitivity. Pseudomonas aeruginosa with multidrug resistance (resistance to three or more antibiotics) was selected to detect the active efflux system of MexAB-OprM type by dilution test of micro broth, and the efflux system and OprD2 were detected by fluorescence quantitative PCR technique. The regulatory gene mexR of high expression MexAB strain was amplified by PCR and the product was sequenced. Results: 1. The most common infection of Pseudomonas aeruginosa was in children and the elderly. The isolation rate of Pseudomonas aeruginosa was the highest in ICU patients in the whole hospital. 2. The drug resistance of Pseudomonas aeruginosa is very severe and has high resistance to most commonly used antibiotics. However, piperacillin / tazobactam and amikacin are still effective antimicrobial agents in the treatment of pseudomonas aeruginosa infection. 3. Proton pump inhibitor CCCP had a great effect on ciprofloxacin. The MIC of 15 strains of Pseudomonas aeruginosa decreased more than 8 times, and the CAZ of 6 strains decreased more than 8 times in the presence of CCCP. In the other two strains, the MIC of only ceftazidime decreased more than 8 times. The antibacterial activity of imipenem was not affected by CCCP. 4. The expression of ceftazidime resistant and sensitive MexAB-OprM active efflux system of Pseudomonas aeruginosa was significantly different (P 0.05), and the expression of MexAB-OprM active efflux system of ciprofloxacin resistance and sensitive two groups of Pseudomonas aeruginosa was statistically significant. P0.05, but resistant to imipenem and sensitive to imipenem, There was no difference in the expression of P0.05 in this type of efflux system. 5. The relative quantitative analysis of mRNA was in good agreement with the results of ceftazidime CCCP inhibition test. 6. The quantitative detection of mRNA of membrane pore protein OprD2 gene showed that the expression of imipenem resistant group was lower than that of sensitive group, and the difference was statistically significant (P0.05). Conclusion: 1. Pseudomonas aeruginosa is an important pathogen of nosocomial infection. 2. The high expression of MexAB-OprM active efflux system in Pseudomonas aeruginosa is an important mechanism of multidrug resistance in Pseudomonas aeruginosa. 3. The high expression of MexAB-OprM type active efflux system was related to the resistance of Pseudomonas aeruginosa to CIP and CAZ, but not to the mechanism of IMP resistance. 4. The decrease of OprD2 expression is an important mechanism of resistance of Pseudomonas aeruginosa to IMP.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R378.991

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本文編號(hào)：1788499