發(fā)布時(shí)間：2018-03-05 07:28

  本文選題：銅綠假單胞菌　切入點(diǎn)：耐藥性　出處：《華中科技大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的和意義 銅綠假單胞菌是引起醫(yī)院感染的重要病原體,多重耐藥的銅綠假單胞菌感染可導(dǎo)致臨床治療困難,病死率增加。而亞胺培南是治療多重耐藥銅綠假單胞菌的有效藥物,隨著它的廣泛使用,對(duì)其耐藥的銅綠假單胞菌日益增多。本文分析亞胺培南耐藥銅綠假單胞菌對(duì)常用抗菌藥物的耐藥性,研究對(duì)亞胺培南的耐藥機(jī)制及所產(chǎn)金屬β-內(nèi)酰胺酶的類型,闡明同濟(jì)醫(yī)院銅綠假單胞菌對(duì)常用抗菌藥物的耐藥性及同源性,為指導(dǎo)臨床合理使用抗菌藥物特別是碳青霉烯類,新藥研究及開(kāi)發(fā),加強(qiáng)感染控制措施,及時(shí)切斷多重耐藥株的克隆傳播和水平傳播,密切監(jiān)測(cè)耐藥株的變遷及其發(fā)展提供科學(xué)依據(jù)。 方法 收集同濟(jì)醫(yī)院2004年1月～9月亞胺培南耐藥銅綠假單胞菌存活株29株,瓊脂稀釋法測(cè)定29株銅綠假單胞菌存活株對(duì)常用的10種抗菌藥物的最低抑菌濃度(minimal inhibitory concentrations,MIC)。通過(guò)雙紙片協(xié)同試驗(yàn)(double-disk synergy testing,DDST)、聚合酶鏈反應(yīng)(polymerase chain reaction ,PCR)、序列分析(sequencing)等方法檢測(cè)金屬β-內(nèi)酰胺酶并分析其類型。紫外分光光度法測(cè)定金屬β-內(nèi)酰胺酶的活性。脈沖場(chǎng)凝膠電泳(pulsed field gelelectrophoresis,PFGE)分析耐藥菌株的同源性。結(jié)果 經(jīng)過(guò)WHONET5.3軟件分析,同濟(jì)醫(yī)院1998年至2004年銅綠假單胞菌對(duì)亞胺培南的耐藥性呈逐年上升趨勢(shì),由1998年7.3%上升至2004年14.3%。同濟(jì)醫(yī)院2004年1～9月共分離銅綠假單胞菌193株,對(duì)頭孢噻肟、慶大霉素和四環(huán)素的耐藥率分別為38.5%、42%和93.7%,其余抗菌藥物的耐藥率均低于20%。對(duì)亞胺培南和美羅培南或頭孢他啶交叉耐藥的銅綠假單胞菌分別為15株(11.0%)和14株(7.3%)。 193株銅綠假單胞菌中,亞胺培南耐藥的銅綠假單胞菌(IMP-R-Pae)共39株。IMP-R-Pae和亞胺培南敏感的銅綠假單胞菌(IMP-S-Pae)對(duì)四環(huán)素的耐藥率均在90%以上;對(duì)哌拉西林/他唑巴坦的耐藥率分別為28.6%和17.9%,差異無(wú)顯著性;對(duì)于其他抗菌藥物的耐藥率,IMP-R-Pae組明顯高于IMP-S-Pae組,且差異呈極顯著性。 藥物敏感性試驗(yàn)結(jié)果顯示,存活的29株亞胺培南耐藥銅綠假單胞菌均為多重耐藥株。頭孢噻肟和頭孢他啶的耐藥率分別為69%和27.6%。氨曲南和阿米卡星的耐藥率均為44.8%,其MIC50和MIC90均為16μg/ml和256μg/ml。亞胺培南和美羅培南的耐藥率分別為93.1%和69%, MIC50和MIC90均為32μg/ml和256μg/ml。29株銅綠假單胞菌中有34.5%(10/29)的菌株對(duì)亞胺培南的MIC32μg/ml,屬于高水平耐藥[11][12]。慶大霉素和阿米卡星的耐藥率分別為82.8%和44.8%,MIC50分別是256μg/ml和16μg/ml, MIC90均為256μg/ml。環(huán)丙沙星、哌拉西林/他唑巴坦和四環(huán)素的耐藥率分別為41.4%、48.3%和100%。 雙紙片協(xié)同試驗(yàn)檢測(cè)29株銅綠假單胞菌中有11株金屬β-內(nèi)酰胺酶陽(yáng)性,陽(yáng)性率為37.9%。金屬β-內(nèi)酰胺酶基因檢測(cè)顯示有2株細(xì)菌產(chǎn)VIM-2型金屬β-內(nèi)酰胺酶(6.9%)。紫外分光光度法檢測(cè)兩株產(chǎn)酶株的酶活性分別為27.35±3.25U,26.58±3.18U。 采用脈沖場(chǎng)凝膠電泳技術(shù)對(duì)29株銅綠假單胞菌進(jìn)行分子分型,PFGE圖譜共分為16個(gè)基因型,A、B、C、D、E、F、G型各有兩株以上,其中D型3株,分別來(lái)自器官移植病房(2株)和神經(jīng)內(nèi)科病房(1株)的不同患者,標(biāo)本采集時(shí)間相近,且均使用過(guò)呼吸機(jī)輔助呼吸,其余類型各1株,兩株產(chǎn)金屬β-內(nèi)酰胺酶菌株不屬于同一基因型,。 結(jié)論 同濟(jì)醫(yī)院2004年1～9月分離的銅綠假單胞菌,除對(duì)頭孢噻肟、慶大霉素和四環(huán)素耐藥率較高,分別為38.5%、42%和93.7%,對(duì)其他抗菌藥物的耐藥率均低于20%。亞胺培南耐藥的銅綠假單胞菌對(duì)于抗菌藥物的耐藥率均高于亞胺培南敏感組,除四環(huán)素和哌拉西林/他唑巴坦外,差異均呈極顯著性(P0.01),且亞胺培南耐藥的銅綠假單胞菌通常表現(xiàn)為多重耐藥。 29株亞胺培南耐藥銅綠假單胞菌,對(duì)頭孢噻肟、慶大霉素、美羅培南和四環(huán)素的耐藥率大于60%;對(duì)環(huán)丙沙星、氨曲南、阿米卡星和哌拉西林/他唑巴坦耐藥率相近,為41%～48%;對(duì)頭孢他啶耐藥率最低,為27.6%。 雙紙片協(xié)同試驗(yàn)產(chǎn)金屬β-內(nèi)酰胺酶菌株陽(yáng)性率為37.9%�；驒z測(cè)只發(fā)現(xiàn)2株產(chǎn)VIM-2型金屬β-內(nèi)酰胺酶。 PFGE分型結(jié)果提示同濟(jì)醫(yī)院亞胺培南耐藥銅綠假單胞菌可能存在小范圍的流行,即可能有耐藥菌株的克隆傳播。同濟(jì)醫(yī)院亞胺培南耐藥銅綠假單胞菌之間尚未發(fā)現(xiàn)滅活酶基因的水平傳播。
[Abstract]:Purpose and significance
Pseudomonas aeruginosa is an important pathogen of nosocomial infection, can lead to difficulties in the clinical treatment of multi drug resistant Pseudomonas aeruginosa infection, the mortality rate increased. Imipenem is an effective drug for the treatment of multi drug resistant Pseudomonas aeruginosa, with its widely used, increasing the drug resistance of Pseudomonas aeruginosa. The commonly used antimicrobial drug resistance of imipenem resistant Pseudomonas aeruginosa strains of this type, the mechanism of imipenem resistance in research and production of metallo beta lactamase, and homology to clarify the drug resistance of Pseudomonas aeruginosa in Tongji Hospital to commonly used antimicrobial drugs, to guide the rational use of clinical antibiotics especially carbapenem class, research and development of new drugs and strengthen infection control measures, cut off the spread of multi drug resistant strains of clonal spread and level in a timely manner, closely monitoring the changes and development of resistant strains and provide the scientific basis According to.
Method
From January 2004 to September in Tongji Hospital of imipenem resistant Pseudomonas strains 29 strains were survival, 29 strains of Pseudomonas aeruginosa strains survived the minimum inhibitory concentration of 10 kinds of commonly used antibiotics by agar dilution method (minimal inhibitory concentrations, MIC). The double disk synergy test (double-disk synergy, testing, DDST), polymerase chain the reaction (polymerase chain reaction, PCR), sequence analysis (sequencing) method lactamases and analysis of its type. The determination of metal beta UV spectrophotometry lactamase activity. Pulsed field gel electrophoresis (pulsed field gelelectrophoresis, PFGE) to analyze the homology of resistant strains.
Through the analysis of WHONET5.3 software, from 1998 to 2004 in Tongji Hospital of Pseudomonas aeruginosa to imipenem increased year by year, up from 7.3% in 1998 to 2004 14.3%. Tongji Hospital in 2004 1~9 months were isolated 193 strains of Pseudomonas aeruginosa, cefotaxime, gentamicin and tetracycline resistance rates were 38.5%, 42% and 93.7%, the drug resistance rates were lower than the 20%. of imipenem and meropenem and ceftazidime interresistant Pseudomonas aeruginosa were 15 strains (11%) and 14 strains (7.3%).
193 strains of Pseudomonas aeruginosa, Pseudomonas aeruginosa resistant to imipenem (IMP-R-Pae and.IMP-R-Pae) a total of 39 strains of imipenem susceptible Pseudomonas aeruginosa (IMP-S-Pae) of tetracycline resistance rate in 90% above; resistant to piperacillin / tazobactam rate were 28.6% and 17.9%, no difference significant; for other antimicrobial drug resistance rate of IMP-R-Pae group was significantly higher than that in IMP-S-Pae group, and the difference was significant.
Drug sensitivity test showed that 29 strains of imipenem resistant Pseudomonas aeruginosa survival were multidrug-resistant. Cefotaxime and ceftazidime resistance rates were 69% and 27.6%. resistant to aztreonam and Amikacin rate were 44.8%, MIC50 and MIC90 were resistant to 16 g/ml and 256 g/ml. imipenem and meropenem were 93.1% and 69%, MIC50 and MIC90 were 34.5% 32 g/ml and 256 g/ml.29 strains of Pseudomonas aeruginosa (10/29) MIC32 g/ml strains to imipenem, resistant to high level resistance to gentamicin and Amikacin [11][12]. respectively 82.8% and 44.8%, respectively, MIC50 256 g/ml and 16 g/ml, 256 g/ml. MIC90 were ciprofloxacin, piperacillin / tazobactam and tetracycline resistance rates were 41.4%, 48.3% and 100%.
Double disk synergy test of 29 strains of Pseudomonas aeruginosa in 11 strains of metallo beta lactamase positive, the positive rate of 37.9%. for the detection of metallo beta lactamase gene showed that 2 strains of VIM-2 producing bacteria of metallo beta lactamase (6.9%). UV spectrophotometry for the determination of enzyme producing strains the activity of the two strains were 27.35 + 3.25U, 26.58 + 3.18U.
The pulsed field gel electrophoresis of 29 strains of Pseudomonas aeruginosa by molecular typing, PFGE map is divided into 16 genotypes, A, B, C, D, E, F, G each have more than two strains, including 3 strains of type D, respectively, from the transplant ward (2 strains) and the Department of Neurology (1 strains) of different patients, the time of sample collection and are similar, use of ventilator assisted breathing, the rest of the type 1 strains, two strains of metallo beta lactamase producing strains do not belong to the same genotype.
conclusion
Pseudomonas aeruginosa isolated from Tongji Hospital in 2004 1~9 months, in addition to cefotaxime, gentamicin and tetracycline resistance rates were 38.5%, 42% and 93.7%, the resistance rates to other antimicrobial agents were lower than 20%. in Pseudomonas aeruginosa imipenem resistant to antibiotics were higher than imipenem sensitive group. In addition to tetracycline and piperacillin / tazobactam, the difference was significantly (P0.01), and Pseudomonas aeruginosa resistant to imipenem usually show multiple drug resistance.
29 strains of imipenem resistant Pseudomonas aeruginosa to gentamicin, cefotaxime, meropenem and tetracycline resistance rate of more than 60%; Ciprofloxacin, aztreonam, Amikacin and piperacillin / tazobactam resistant rate was 41% ~ 48%; similar to ceftazidime resistant rate of the low 27.6%.
The positive rate of metallo beta lactamase producing strain was detected by 37.9%. gene and only 2 strains of VIM-2 type metallo beta lactamase were found in the double paper synergistic test.
The results of PFGE typing suggested that there may be a small epidemic of imipenem resistant Pseudomonas aeruginosa in Tongji Hospital, that is, there may be a clone propagation of drug-resistant strains. No level of inactivated enzyme gene has been found between imipenem resistant Pseudomonas aeruginosa in Tongji Hospital.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R181.3

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