本文選題：銅綠假單胞菌 + 多藥耐藥　； 參考：《中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院》2012年碩士論文

【摘要】：目的：1、研究高齡患者呼吸機(jī)相關(guān)性肺炎的病原菌分布及耐藥情況。2、探討引起呼吸機(jī)相關(guān)性肺炎的銅綠假單胞菌耐藥機(jī)制。3、對(duì)多藥耐藥銅綠假單胞菌進(jìn)行體外聯(lián)合藥敏試驗(yàn)，，為臨床合理用藥提供依據(jù)。 方法： 1、收集2007年1月～2011年6月診斷為VAP的高齡患者123例，研究其病原菌分布及耐藥情況。 2、將兩周之內(nèi)，同一患者且對(duì)某一類抗菌藥物藥敏有變化的兩株銅綠假單胞菌配為一對(duì)，其中碳青霉烯類20對(duì)，喹諾酮類22對(duì)，氨基糖苷類15對(duì)，多粘菌素B10對(duì)。收集來(lái)自30個(gè)高齡VAP患者的PA122株。 3、瓊脂稀釋法檢測(cè)PA對(duì)14種抗生素的MIC、外排泵表型。PFGE法檢測(cè)基因同源性。雙紙片協(xié)同法檢測(cè)金屬酶表型。PCR法檢測(cè)耐藥基因。結(jié)晶紫染色法檢測(cè)細(xì)菌生物被膜形成能力。掃描電鏡觀察生物被膜。棋盤法設(shè)計(jì)，瓊脂稀釋法行體外聯(lián)合藥敏。 結(jié)果： 1、123例高齡老年VAP患者共檢出細(xì)菌409株，其中革蘭氏陰性菌355株，革蘭氏陽(yáng)性菌54株，革蘭陰性菌中銅綠假單胞菌24.44%、鮑曼不動(dòng)桿菌17.11%、嗜麥芽窄食單胞14.67%、洋蔥伯克霍爾德菌7.58%;革蘭陽(yáng)性菌中以金黃色葡萄球菌9.05%為主。銅綠假單胞菌對(duì)常用抗菌藥物的耐藥性較高。 2、所收集的配對(duì)PA菌株共來(lái)自30個(gè)患者，其中70%的患者痰標(biāo)本分離的配對(duì)菌株經(jīng)PFGE證明為同一基因來(lái)源，30%的患者痰標(biāo)本分離的配對(duì)菌株經(jīng)PFGE證明為不同基因來(lái)源。 3、4個(gè)患者共8株菌擴(kuò)增出OXA-1基因，3個(gè)患者共6株菌擴(kuò)增出OXA-10基因，僅一個(gè)患者1株菌擴(kuò)增出GES基因，13個(gè)患者共33株菌擴(kuò)增出OprD2基因，余菌株為OprD2基因缺失或部分缺失菌株。未擴(kuò)增出KPC、GIM、OXA-2、VIM及IMP耐藥基因。同一患者來(lái)源且經(jīng)PFGE證明為同一基因來(lái)源的菌株中，PCR法檢測(cè)其耐藥基因型如OXA-1、OXA-10、GES、OprD2并不完全一致。僅有一個(gè)患者分離的共4株菌金屬酶表型陽(yáng)性。5個(gè)患者共9株菌外排泵表型陽(yáng)性。所有菌株均有不同程度的生物膜形成能力。 4、各組藥物聯(lián)用后，除TZP和CIP聯(lián)合主要表現(xiàn)為無(wú)關(guān)作用外，其他組合均主要表現(xiàn)為協(xié)同和相加作用，濃度-累積抑菌百分率曲線均左移。結(jié)論：（1）PA在高齡VAP患者中分離率占首位，且高度耐藥。（2）70%老年P(guān)A-VAP患者自身不同時(shí)間點(diǎn)所分離的PA為同一基因來(lái)源。（3）PA的高度耐藥為以產(chǎn)生物膜為主的多種機(jī)制共同作用結(jié)果。（4）MDRP可選聯(lián)合用藥方案。
[Abstract]:Objective to study the distribution and drug resistance of pathogenic bacteria in ventilator-associated pneumonia (VAP) in elderly patients, and to explore the mechanism of resistance of Pseudomonas aeruginosa to multidrug resistant Pseudomonas aeruginosa in vitro, and to investigate the mechanism of drug resistance of Pseudomonas aeruginosa, and to study the drug resistance of Pseudomonas aeruginosa in vitro. To provide the basis for clinical rational use of drugs. Methods: 1. 123 elderly patients diagnosed as VAP from January 2007 to June 2011 were collected to study the distribution of pathogenic bacteria and drug resistance. Two strains of Pseudomonas aeruginosa were matched in the same patient, including 20 pairs of carbapenems, 22 pairs of quinolones, 15 pairs of aminoglycosides and 10 pairs of polymyxin B _ (10). PA122 strains from 30 old patients with VAP were collected. The MICs of PA to 14 antibiotics were detected by Agar dilution method. The homology of gene was detected by efflux pump phenotype. PFGE method. Detection of metallozyme phenotypes by double disk synergy. PCR was used to detect drug resistance genes. The ability of bacterial biofilm formation was detected by crystal violet staining. The biofilm was observed by scanning electron microscope. Chessboard design, Agar dilution method combined with in vitro drug sensitivity. Results: a total of 409 strains of bacteria were detected in 1123 elderly patients with VAP, of which 355 were Gram-negative bacteria and 54 were Gram-positive bacteria. Pseudomonas aeruginosa 24.44m, Acinetobacter baumannii 17.11m, maltophilia 14.677m, Bacillus cepacia 7.58m, Gram-positive bacteria 9.05%. Pseudomonas aeruginosa has higher resistance to common antimicrobial agents. 2. Pairing PA strains were collected from 30 patients. Among them, 70% of the matched strains isolated from sputum samples from patients were identified as the same gene source by PFGE, and 30% of them were identified as different gene sources by PFGE. OXA-1 was amplified from 8 strains from 34 patients. OXA-10 gene was amplified from 6 strains of 3 patients. GES gene was amplified from one strain of one patient and OprD2 gene was amplified from 33 strains of 13 patients. The remaining strains were OprD2 gene deletion or partial deletion. KPC-GIMA OXA-2 vim and IMP resistance genes were not amplified. The genotypes of OXA-1, OXA-10, GESS-OprD2, which were identified by PFGE as the same gene source, were not identical with each other by polymerase chain reaction (PCR). Only 4 strains were positive for metallozyme phenotype and 9 strains were positive for efflux pump phenotype in 5 patients. All the strains had different biofilm forming ability. 4. After combined use of TZP and CIP, the combination of TZP and CIP were mainly synergistic and additive. The curves of concentration-cumulative bacteriostatic percentage were all shifted to the left. Conclusion: (1) the isolation rate of PA in the elderly patients with VAP is the highest. And high drug resistance. (2) PA isolated from 70% old PA-VAP patients at different time points was the same gene source. (3) the high resistance of PA was the result of multiple mechanisms of biofilm production. (4) MDRP could be used in combination.
【學(xué)位授予單位】：中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R563.1

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