本文選題：銅綠假單胞菌 + 多藥耐藥��； 參考：《中國人民解放軍軍醫(yī)進修學院》2012年碩士論文

【摘要】：目的：1、研究高齡患者呼吸機相關性肺炎的病原菌分布及耐藥情況。2、探討引起呼吸機相關性肺炎的銅綠假單胞菌耐藥機制。3、對多藥耐藥銅綠假單胞菌進行體外聯(lián)合藥敏試驗，，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，同一患者且對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，其中碳青霉烯類20對，喹諾酮類22對，氨基糖苷類15對，多粘菌素B10對。收集來自30個高齡VAP患者的PA122株。 3、瓊脂稀釋法檢測PA對14種抗生素的MIC、外排泵表型。PFGE法檢測基因同源性。雙紙片協(xié)同法檢測金屬酶表型。PCR法檢測耐藥基因。結晶紫染色法檢測細菌生物被膜形成能力。掃描電鏡觀察生物被膜。棋盤法設計，瓊脂稀釋法行體外聯(lián)合藥敏。 結果： 1、123例高齡老年VAP患者共檢出細菌409株，其中革蘭氏陰性菌355株，革蘭氏陽性菌54株，革蘭陰性菌中銅綠假單胞菌24.44%、鮑曼不動桿菌17.11%、嗜麥芽窄食單胞14.67%、洋蔥伯克霍爾德菌7.58%;革蘭陽性菌中以金黃色葡萄球菌9.05%為主。銅綠假單胞菌對常用抗菌藥物的耐藥性較高。 2、所收集的配對PA菌株共來自30個患者，其中70%的患者痰標本分離的配對菌株經(jīng)PFGE證明為同一基因來源，30%的患者痰標本分離的配對菌株經(jīng)PFGE證明為不同基因來源。 3、4個患者共8株菌擴增出OXA-1基因，3個患者共6株菌擴增出OXA-10基因，僅一個患者1株菌擴增出GES基因，13個患者共33株菌擴增出OprD2基因，余菌株為OprD2基因缺失或部分缺失菌株。未擴增出KPC、GIM、OXA-2、VIM及IMP耐藥基因。同一患者來源且經(jīng)PFGE證明為同一基因來源的菌株中，PCR法檢測其耐藥基因型如OXA-1、OXA-10、GES、OprD2并不完全一致。僅有一個患者分離的共4株菌金屬酶表型陽性。5個患者共9株菌外排泵表型陽性。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)為無關作用外，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PA-VAP患者自身不同時間點所分離的PA為同一基因來源。（3）PA的高度耐藥為以產(chǎn)生物膜為主的多種機制共同作用結果。（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.
【學位授予單位】：中國人民解放軍軍醫(yī)進修學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R563.1

【參考文獻】

相關期刊論文 前10條

1 左聯(lián);綠膿桿菌外膜通透性與耐藥性的關系[J];國外醫(yī)學.藥學分冊;1997年03期

2 張永,唐英春;病原菌質子驅動型外排泵分子機制研究進展[J];國外醫(yī)藥(抗生素分冊);2004年01期

3 楊菊;劉彬;瞿秋;;硫酸依替米星在老年人感染性疾病應用的安全性分析[J];湖南中醫(yī)藥大學學報;2010年02期

4 王麗麗;徐建國;;脈沖場凝膠電泳技術(PFGE)在分子分型中的應用現(xiàn)狀[J];疾病監(jiān)測;2006年05期

5 王衛(wèi)萍;邵海楓;韓麗麗;王錦娜;史利寧;張小衛(wèi);;銅綠假單胞菌對亞胺培南耐藥機制的研究[J];醫(yī)學研究生學報;2006年11期

6 王潔;邵海楓;;腸桿菌科質粒介導喹諾酮類抗菌藥耐藥的研究[J];醫(yī)學研究生學報;2008年01期

7 磨國鑫;管希周;崔俊昌;佘丹陽;陳良安;;65株鮑曼不動桿菌的臨床分布和耐藥特征分析[J];軍醫(yī)進修學院學報;2011年07期

8 梅亞寧;文怡;陳友華;劉根焰;張巧娣;趙旺勝;;美羅培南和亞胺培南-西司他丁對100株革蘭陰性桿菌體外抗菌活性比較[J];中國感染與化療雜志;2006年01期

9 汪復;朱德妹;胡付品;阮斐怡;倪語星;孫景勇;徐英春;張小江;胡云健;艾效曼;俞云松;楊青;孫自鏞;李麗;賈蓓;黃文祥;卓超;蘇丹虹;魏蓮花;吳玲;張朝霞;季萍;王傳清;薛建昌;張泓;李萬華;徐元宏;沈繼錄;單斌;杜艷;;2009年中國CHINET細菌耐藥性監(jiān)測[J];中國感染與化療雜志;2010年05期

10 王傳馥;21世紀老年醫(yī)學展望[J];老年醫(yī)學與保健;2001年01期

本文編號：2064843