本文關(guān)鍵詞：耐環(huán)丙沙星鮑曼不動(dòng)桿菌主動(dòng)外排機(jī)制的研究及耐藥逆轉(zhuǎn)初步探討　出處：《中南大學(xué)》2009年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 鮑曼不動(dòng)桿菌 環(huán)丙沙星 主動(dòng)外排泵 抑制劑

【摘要】： 目的了解長(zhǎng)沙地區(qū)鮑曼不動(dòng)桿菌對(duì)環(huán)丙沙星的耐藥現(xiàn)狀,為臨床治療鮑曼不動(dòng)桿菌感染、合理應(yīng)用喹諾酮類(lèi)抗生素提供指導(dǎo);研究鮑曼不動(dòng)桿菌對(duì)喹諾酮類(lèi)藥物的耐藥性與主動(dòng)外排機(jī)制的關(guān)系;初步探討鮑曼不動(dòng)桿菌對(duì)喹諾酮類(lèi)藥物的耐藥逆轉(zhuǎn)機(jī)制,尋求恢復(fù)其對(duì)喹諾酮類(lèi)藥物敏感性的方法。 方法(1)微量稀釋法檢測(cè)56株鮑曼不動(dòng)桿菌對(duì)環(huán)丙沙星的敏感性。(2)采用直接熒光法篩選出胞內(nèi)環(huán)丙沙星聚集量明顯差異的菌株作為耐藥組,并測(cè)定敏感組和耐藥組在有無(wú)外排泵抑制劑情況下,鮑曼不動(dòng)桿菌對(duì)環(huán)丙沙星的吸收和積累情況。(3)用逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)方法檢測(cè)主動(dòng)外排泵基因adeABC的表達(dá)水平。(4)擴(kuò)增adeABC自身啟動(dòng)子區(qū)域片段基因并測(cè)序。(5)逆轉(zhuǎn)實(shí)驗(yàn)觀察三種抑制劑對(duì)外排泵的抑制作用。 結(jié)果(1)56株鮑曼不動(dòng)桿菌中,對(duì)環(huán)丙沙星耐藥的有48株(MIC＞4 mg/l);對(duì)環(huán)丙沙星中介有0株(1 mg/l≤MIC≤4 mg/l);對(duì)環(huán)丙沙星敏感的有8株(MIC＜1 mg/l)。(2)耐環(huán)丙沙星的所有菌株中,挑選胞內(nèi)環(huán)丙沙星藥物濃度累積差異最大的6株作為耐藥組(R組),與敏感組(S組)相比較:無(wú)NaN_3時(shí),S組的累積量(138.01±0.251 ng/ml)明顯高于R組(62.15±0.743 ng/ml)(P＜0.01);加入NaN_3后,R組的累積量明顯上升(114.78±0.415 ng/ml),與無(wú)NaN_3的S組(138.01±0.251 ng/ml)無(wú)差異(P＞0.05),但是與無(wú)NaN_3的R組(62.15±0.743 ng/ml)有明顯差異(P＜0.05);加NaN_3后的S組(138.27±0.177 ng/ml)與未加NaN_3時(shí)(138.01±0.251 ng/ml)無(wú)明顯差異(P＞0.05)。(3)耐藥組主動(dòng)外排基因adeB的表達(dá)水平(2.372±0.032)明顯高于敏感組(1.654±0.040)(P＜0.01)。(4)adeABC自身啟動(dòng)子區(qū)域沒(méi)有發(fā)現(xiàn)有意義突變。(5)分別在環(huán)丙沙星中加入外排泵抑制劑NaN_3、MC-207110或利血平后,臨床分離敏感株MIC值變化不大;臨床分離耐藥株MIC均有所下降,且MIC值下降至原值1/4的株數(shù)(即外排陽(yáng)性株)分別為6株、6株和2株。 結(jié)論:(1)長(zhǎng)沙地區(qū)鮑曼不動(dòng)桿菌對(duì)環(huán)丙沙星的耐藥率高達(dá)85.7%。(2)adeABC基因的過(guò)度表達(dá)是鮑曼不動(dòng)桿菌對(duì)環(huán)丙沙星耐藥的重要原因之一。(3)在對(duì)環(huán)丙沙星耐藥的鮑曼不動(dòng)桿菌中,adeB自身啟動(dòng)子區(qū)域未見(jiàn)有意義突變。(4)使用外排泵抑制劑MC-207110和NaN_3后耐藥株對(duì)環(huán)丙沙星的敏感性有顯著的恢復(fù);利血平對(duì)外排泵adeABC的抑制作用不如前兩者明顯。
[Abstract]:Objective to understand the Changsha area Bauman real status of resistance to ciprofloxacin for the treatment of Bauman coli, Acinetobacter infection, rational use of quinolones to provide guidance; the study of relationship between Bauman real resistance and active bacillus of quinolone efflux mechanism; preliminary study of Bauman Acinetobacter quinolone resistance reversal mechanism, seeking to restore the method of Quinolone drug sensitivity.
Methods (1) detection of trace dilution method in 56 strains of Acinetobacter strains to ciprofloxacin susceptibility to Bauman. (2) screened the intracellular accumulation of ciprofloxacin significantly different resistant strains as group by immunofluorescence assay and determination of sensitive group and resistant group in the presence of efflux pump inhibitors under Bauman real and accumulation the absorption of ciprofloxacin. Coli (3) by reverse transcription polymerase chain reaction (RT-PCR) pump gene expression adeABC method for detection of active efflux. (4) the amplification of adeABC promoter region gene fragment and sequencing. (5) observed the reversal of inhibition of three kinds of inhibitors efflux pump.
緇撴灉(1)56鏍矋鏇間笉鍔ㄦ潌鑿屼腑,瀵圭幆涓欐矙鏄熻，

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