本文關(guān)鍵詞： 多重耐藥鮑曼不動(dòng)桿菌 耐藥基因 聚合酶鏈反應(yīng) 脈沖場(chǎng)凝膠電泳　出處：《四川醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：鮑曼不動(dòng)桿菌是一類非發(fā)酵的革蘭陰性桿菌,為條件致病菌。廣泛存在于水、土壤、醫(yī)院環(huán)境和人體皮膚表面,是醫(yī)院感染的重要致病菌之一,主要引起獲得性肺炎特別是呼吸機(jī)相關(guān)性肺炎、菌血癥、尿路感染、繼發(fā)性腦膜炎等。隨著廣譜抗菌藥物和免疫抑制劑大量使用,對(duì)常用的β-內(nèi)酰胺類、氨基糖苷類、氟喹諾酮類等抗菌藥物同時(shí)耐藥的多重耐藥鮑曼不動(dòng)桿菌分離率明顯增多,并在醫(yī)院內(nèi)廣泛流行,給臨床抗感染治療帶來(lái)極大的麻煩,同時(shí)對(duì)醫(yī)院感染控制提出了新的挑戰(zhàn)。對(duì)多重耐藥鮑曼不動(dòng)桿菌(Multi-drug resistant Acinetobacter baumannii,MDRAB)的藥敏試驗(yàn)和醫(yī)院科室分布情況進(jìn)行統(tǒng)計(jì)和分析,并檢測(cè)相關(guān)耐藥基因和對(duì)菌株進(jìn)行同源性分析,從而指導(dǎo)醫(yī)生合理使用抗菌藥物,探討我院多重耐藥鮑曼不動(dòng)桿菌產(chǎn)生的耐藥機(jī)制,為我院預(yù)防與控制院內(nèi)感染提供理論依據(jù),給流行病學(xué)研究奠定基礎(chǔ)。本研究收集四川醫(yī)科大學(xué)附屬第一醫(yī)院2013年9月至2014年9月臨床分離的62株非重復(fù)多重耐藥鮑曼不動(dòng)桿菌。利用MicroScan WalkAway96 Plus全自動(dòng)微生物鑒定/藥敏測(cè)試系統(tǒng)進(jìn)行鑒定、驗(yàn)證,瓊脂稀釋法檢測(cè)菌株對(duì)23種抗生素的最低抑菌濃度(MIC),利用聚合酶鏈反應(yīng)(Polymerase Chain Reaction,PCR)檢測(cè)菌株的6種相關(guān)耐藥基因(TEM、SHV、VIM、IMP、OXA-23、OXA-24)的攜帶情況,將擴(kuò)增所得的陽(yáng)性產(chǎn)物送上海鉑尚生物技術(shù)有限公司測(cè)序,將測(cè)序所得序列和GenBank數(shù)據(jù)庫(kù)進(jìn)行對(duì)比分析,并采用脈沖場(chǎng)凝膠電泳(Pulsed Field Gel Electrophoresis,PFGE)對(duì)62株多重耐藥鮑曼不動(dòng)桿菌進(jìn)行分子分型并分析其同源性。結(jié)果顯示:62株多重耐藥鮑曼不動(dòng)桿菌主要分離于痰液,其中40株分離于痰液標(biāo)本,11株分離于分泌物,其余為分離于血液標(biāo)本4株、中段尿標(biāo)本3株、腦脊液標(biāo)本2株和胸水標(biāo)本2株。臨床科室分布分別是重癥監(jiān)護(hù)室24株,呼吸內(nèi)科10株,神經(jīng)外科8株,余下菌株來(lái)源于其他臨床科室。62株多重耐藥鮑曼不動(dòng)桿菌對(duì)青霉素類的耐藥率均為100%;對(duì)頭孢菌素中頭孢曲松鈉耐藥率為96.8%,其余均為100%。對(duì)氨芐西林/舒巴坦的耐藥率為80.6%,對(duì)碳青霉烯類抗生素中亞胺培南和美羅培南的耐藥率為90.3%和88.7%,對(duì)氨基糖苷類抗生素中慶大霉素、阿米卡星、妥布霉素的耐藥率分別為93.5%、80.6%和53.2%,對(duì)喹諾酮類抗生素中環(huán)丙沙星、左氧氟沙星、加替沙星的耐藥率分別為100%、85.5%和64.5%,對(duì)多粘菌素和多粘菌素b的耐藥率為12.9%和11.3%。對(duì)四環(huán)素、米諾環(huán)素、替加環(huán)素的耐藥率分別為90.3%、17.7%、3.2%。對(duì)磺胺甲惡唑和利福平的耐藥率為69.4%和92%。6種耐藥基因共檢出4種,其中tem陽(yáng)性有52株,陽(yáng)性率為83.9%,vim陽(yáng)性有53株,陽(yáng)性率為85.5%,imp陽(yáng)性有41株,陽(yáng)性率為66.1%,oxa-23陽(yáng)性有58株,陽(yáng)性率為93.5%。4種耐藥基因陽(yáng)性擴(kuò)增產(chǎn)物經(jīng)測(cè)序后與genbank中已提交的相應(yīng)基因序列的同源性為98%-100%。pfge將62株多重耐藥鮑曼不動(dòng)桿菌分為4型,a型和b型為主要克隆株。a型包括4個(gè)亞型共32株,b型有2個(gè)亞型共19株,c克隆有8株。d克隆有3株。以上研究表明我院分離的多重耐藥鮑曼不動(dòng)桿菌主要分布在呼吸內(nèi)科、重癥監(jiān)護(hù)室、神經(jīng)外科,呼吸道感染在感染病例中為主,與接受各種侵襲性操作有關(guān)。多重耐藥鮑曼不動(dòng)桿菌的耐藥情況十分嚴(yán)峻,62株耐藥菌株對(duì)23種抗生素的耐藥水平高于全國(guó)細(xì)菌耐藥監(jiān)測(cè)網(wǎng)的水平。oxa-23在62株多重耐藥鮑曼不動(dòng)桿菌中的攜帶率最高,為93.5%。并且攜帶vim、tem、imp菌株也高達(dá)85.5%、83.9%、66.1%,說(shuō)明攜帶多種耐藥基因是在多重耐藥鮑曼不動(dòng)桿菌的中十分普遍。我院鮑曼不動(dòng)桿菌pfge分型分為4型,以a型為主,共有32株占51.6%,主要在重癥監(jiān)護(hù)室、呼吸內(nèi)科、神經(jīng)外科、骨科、血液科5個(gè)病區(qū)流行,其次為B型,主要集中在呼吸內(nèi)科、重癥監(jiān)護(hù)室和兒科。多重耐藥鮑曼不動(dòng)桿菌在我院各臨床科室廣泛分布并流行傳播,克隆株在醫(yī)院內(nèi)部和醫(yī)院之間克隆蔓延已成為全球性的問(wèn)題,成為耐藥菌株逐年增加的一個(gè)重要因素。因此,應(yīng)對(duì)醫(yī)院感染控制加大力度,制定更加嚴(yán)格的院感制度,慎重合理使用抗菌藥物,防止多重耐藥鮑曼不動(dòng)桿菌的爆發(fā)流行。
[Abstract]:Bauman Acinetobacter is a kind of non fermentative gram negative bacteria are opportunistic bacteria. Widely exist in water, soil, hospital environment and human skin is an important pathogen of nosocomial infection, the main cause of pneumonia especially ventilator-associated pneumonia, bacteremia, urinary tract infection, secondary meningitis etc. with the use of antibacterial drugs and immunosuppressive agents of common broad-spectrum beta lactams, aminoglycosides, multi drug resistant Bauman fluoroquinolones and other antimicrobial agents and drug resistance of Acinetobacter isolated rate increased significantly, and widely popular in the hospital for clinical anti infection treatment brought great trouble to the hospital at the same time the infection control has brought new challenges. For multi drug resistant Acinetobacter Bauman (Multi-drug resistant Acinetobacter baumannii, MDRAB) of the drug sensitivity test and the distribution of hospital departments of statistics and Analysis and detection of drug resistance related genes and to analyze the homology of the strains, so as to guide the rational use of antimicrobial drugs doctors in our hospital, to explore multidrug resistance mechanisms of Bauman, provide a theoretical basis for the prevention and control of hospital infection in our hospital, to lay the foundation for epidemiological research. This study collects the Sichuan Medical University the first hospital from September 2013 to September 2014 a total of 62 strains of non repetitive multi drug resistant Acinetobacter Bauman. Confirmed by MicroScan WalkAway96 Plus automatic microorganism identification / drug sensitivity test system were identified, the minimum inhibitory concentration of agar dilution method to detect strains to 23 antibiotics (MIC), using polymerase chain reaction (Polymerase Chain Reaction. PCR) 6 kinds of drug resistance related gene detection strains (TEM, SHV, VIM, IMP, OXA-23, OXA-24) the carrying status of positive products of the amplified platinum is sent to Shanghai students Material Technology Co. Ltd. will be sequenced, sequence and GenBank database were analyzed by pulsed field gel electrophoresis (Pulsed Field Gel Electrophoresis, PFGE) of 62 strains of multidrug-resistant Acinetobacter Bauman for molecular typing and homology analysis. The results showed that 62 strains of multidrug-resistant Acinetobacter isolated from major Bauman in the sputum, 40 of which were isolated from sputum specimens, 11 strains were isolated from the rest of secretions, is separated from the blood samples of 4 strains, 3 strains of urine, cerebrospinal fluid specimens of 2 strains and 2 strains of pleural effusion specimens. The distribution of clinical departments were ICU 24 strains, 10 strains of respiratory medicine, Department of Neurosurgery, 8 strains. The remaining strains from other clinical departments.62 strains of multi resistant Acinetobacter Bauman resistant to penicillin rate was 100%; the cephalosporin ceftriaxone resistance rate was 96.8%, the rest are 100%. to ampicillin / sulbactam The resistant rate was 80.6%, the resistance to imipenem in carbapenems imipenem and meropenem were 90.3% and 88.7%, the aminoglycoside antibiotic gentamicin, Amikacin, tobramycin resistance rates were 93.5%, 80.6% and 53.2% of the quinolones antibiotics ciprofloxacin, levofloxacin, gatifloxacin resistance rates were 100%, 85.5% and 64.5% of polymyxin B and polymyxin resistance rate of 12.9% and 11.3%. of tetracycline, minocycline, tigecycline resistance rates were 90.3%, 17.7%, 3.2%. resistant to sulfamethoxazole and rifampin were 69.4% and 92%.6 resistant genes were detected in 4, among them there are 52 TEM positive strains, the positive rate was 83.9%, VIM positive 53 strains, the positive rate was 85.5%, imp positive 41 strains, the positive rate was 66.1%, OXA-23 positive 58 strains, the positive rate of 93.5%.4 gene positive PCR products were sequenced and Gen The homology of the corresponding gene sequence has been submitted to the bank in the 98%-100%.pfge of 62 strains of multi resistant Acinetobacter Bauman is divided into 4 types, type A and B as the main clone. Type A includes 4 subtypes of 32 strains, 2 B subtype 19 strains, 8 strains of C clone.D clone 3 the study indicated that multi drug resistant strains. Bauman in our hospital isolates of Acinetobacter are mainly distributed in the Department of respiratory medicine, Department of Neurosurgery, ICU, mainly in case of infection of respiratory tract infection, and accept a variety of invasive operation. Multidrug resistant Bauman real drug resistance bacteria is very serious, the level of resistance of 62 strains of resistant strains to 23 antibiotics is higher than that of mohnarin level.Oxa-23 in 62 strains of multidrug-resistant Acinetobacter Bauman in carrying the highest rate, carrying VIM, 93.5%. and TEM, imp was as high as 85.5%, 83.9%, 66.1%, that carry multiple resistance genes in multiple Bauman resistant Acinetobacter is common in our hospital. Bauman Acinetobacter PFGE divided into 4 types, with a total of 32 strains, accounting for 51.6%, mainly in the ICU, respiratory medicine, Department of Neurosurgery, Department of orthopedics, Department of Hematology, 5 wards pop, followed by B, mainly concentrated in the Department of respiratory medicine ICU, and pediatrics. Multi drug resistant Acinetobacter Bauman in all clinical departments in our hospital are widely distributed and spreading, clones between the hospital and hospital internal clone spread has become a global problem, become drug resistant strains increased year by year one of the most important factors. Therefore, to deal with the hospital infection control efforts to develop more strict hospital system, carefully rational use of antimicrobial drugs to prevent multidrug resistant Bauman real epidemic coli outbreak.

【學(xué)位授予單位】：四川醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 趙旺勝;江淑芳;顧兵;吳姍姍;劉根焰;張巧娣;吳海瑋;;南京地區(qū)鮑曼不動(dòng)桿菌喹諾酮類藥物耐藥基因突變的研究[J];臨床檢驗(yàn)雜志;2007年02期

2 邢麗丹;糜祖煌;徐鑫鑫;汪汀;田莎莎;蘇兆亮;許化溪;;多重耐藥鮑曼不動(dòng)桿菌中β內(nèi)酰胺酶基因的檢測(cè)[J];中國(guó)感染與化療雜志;2014年01期

3 應(yīng)春妹;翁文浩;;鮑曼不動(dòng)桿菌多重耐藥機(jī)制研究進(jìn)展[J];檢驗(yàn)醫(yī)學(xué);2007年02期

4 羅柳林;應(yīng)春妹;倪培華;葉楊芹;汪雅萍;張灝e，

本文編號(hào)：1529627