【摘要】：目的:了解我院242株臨床分離鮑曼不動(dòng)桿菌感染患者的臨床特征,檢測(cè)其對(duì)常見(jiàn)抗菌藥物敏感性,檢測(cè)外排泵基因分布及部分外排泵基因表達(dá)水平,初探鮑曼不動(dòng)桿菌耐替加環(huán)素的機(jī)制。方法:收集川北醫(yī)學(xué)院附屬醫(yī)院2015年9月至2016年9月培養(yǎng)到的鮑曼不動(dòng)桿菌株和相關(guān)臨床資料,用K-B法對(duì)其進(jìn)行體外藥敏試驗(yàn),并用PCR方法對(duì)adeB,adeG,adeJ,adeR,ade S,adeE,abeM,tetA,tetB,abeS等外排泵基因進(jìn)行檢測(cè),對(duì)同時(shí)檢出adeB,adeG,adeJ,abeM的菌株根據(jù)藥敏試驗(yàn)分為對(duì)替加環(huán)素敏感組和耐藥組,利用qPCR方法檢測(cè)ade B,adeG,adeJ,abeM表達(dá)量并進(jìn)行比較。結(jié)果:1.242株臨床分離鮑曼不動(dòng)桿菌標(biāo)本來(lái)源于痰液最多,達(dá)181株(占74.79%),其次是膿液41株(16.94%)�？剖曳植记叭环謩e為:ICU(45.04%),神經(jīng)外科(19.01%)和呼吸內(nèi)科(18.18%)。顯示男性138例,女性104例;≥70歲的有157例,50~70歲的64例;合并顱腦疾病最多,有65例(26.86%),合并COPD、糖尿病、心血管疾病分別有56例(23.14%)、46例(19.00%)、42例(17.36%)。合并器官衰竭情況:呼吸衰竭101例(41.73%),心功能衰竭39例(16.11%),腎功能衰竭15例(6.20%);抗菌藥物使用情況:II聯(lián)用藥147例(60.74%),I聯(lián)用藥62例(25.62%),III聯(lián)及以上用藥33例(13.64%);住院期間使用過(guò)3種及以上抗菌藥物的有149例(61.57%),使用過(guò)碳青霉烯類類抗生素的有87例(35.95%);有侵襲性操作者占86.78%;人工氣道或機(jī)械通氣者占62%;預(yù)后:住院期間無(wú)明顯好轉(zhuǎn)或死亡的有92例(38.02%);平均住院天數(shù)46.2天。2.我院臨床分離的242株鮑曼不動(dòng)桿菌體外藥敏試驗(yàn)顯示:鮑曼不動(dòng)桿菌對(duì)頭孢哌酮-舒巴坦、美羅培南、亞胺培南、替加環(huán)素耐藥率50%,分別為45.04%、29.75%、27.27%、3.31%;對(duì)頭孢吡肟、頭孢曲松、頭孢噻肟、頭孢他啶耐藥率分別為75.21%、73.55%、71.07%、66.53%;對(duì)鏈霉素、慶大霉素、氨芐西林/舒巴坦、哌拉西林/他唑巴坦、環(huán)丙沙星、左氧氟沙星、阿米卡星耐藥率分別為:87.60%、83.06%、78.10%、77.69%、77.69%、66.12%、65.29%。3.我院242株鮑曼不動(dòng)桿菌外排泵基因adeb,adeg,adej,ader,ades,adee,abem,teta,tetb,abes檢出率分別為:81.82%、85.95%、83.47%、81.00%、78.93%、0%、80.58%、21.49%、46.28%、100%。對(duì)同時(shí)檢測(cè)到adeb、adeg、adej、abem基因的菌株以對(duì)替加環(huán)素耐藥與否分為敏感組和耐藥組,檢測(cè)其adeb、adeg、adej、abem的表達(dá)量,并對(duì)其進(jìn)行比較,結(jié)果顯示:adeb、adeg、adej、abem的表達(dá)量在耐藥組:敏感組分別為:21倍、1倍、5倍、1倍。結(jié)論:1.我院鮑曼不動(dòng)桿菌菌株分布主要在icu,其次是神經(jīng)外科、呼吸內(nèi)科,主要來(lái)源于痰液,感染患者年齡偏大,大部分在70歲以上,合并基礎(chǔ)疾病多且病情重,住院期間抗菌藥物使用種類多,聯(lián)用現(xiàn)象常見(jiàn),以肺部感染居多,容易發(fā)生呼吸衰竭,侵襲性操作或手術(shù)可能為鮑曼不動(dòng)桿菌感染的危險(xiǎn)因素。2.我院臨床分離鮑曼不動(dòng)桿菌對(duì)頭孢吡肟、頭孢曲松、頭孢噻肟、頭孢他啶耐藥率高,對(duì)頭孢哌酮-舒巴坦耐藥率高于美羅培南、亞胺培南等碳青霉烯類抗菌藥物,對(duì)替加環(huán)素耐藥率低,碳青霉烯類抗菌藥物、替加環(huán)素可以作為我院臨床治療鮑曼不動(dòng)桿菌感染的首選藥物。3.此次試驗(yàn)鮑曼不動(dòng)桿菌中RND(adeABC、adeFGH、adeIJK)外排泵基因檢出率普遍高,耐藥組檢出率高于敏感組,故鮑曼不動(dòng)桿菌對(duì)抗菌藥物的耐藥可能主要由RND外排泵介導(dǎo)有關(guān);adeE檢出率為0,證明我院鮑曼不動(dòng)桿菌可能不存在adeE外排泵基因流行現(xiàn)象。4.外排泵adeABC、adeIJK的高表達(dá)介導(dǎo)我院鮑曼不動(dòng)桿菌對(duì)替加環(huán)素耐藥,而RND外排泵家族中的adeFGH和abeM(即MATE外排泵系統(tǒng))是否介導(dǎo)我院鮑曼不動(dòng)桿菌對(duì)替加環(huán)素耐藥需擴(kuò)大樣本進(jìn)一步研究。
[Abstract]:Objective: To investigate the clinical characteristics of 242 clinical isolates of Baumannii infection in our hospital, to detect the susceptibility to common antimicrobial agents, to detect the distribution of efflux pump gene and the expression level of partial efflux pump gene. Methods: The drug sensitivity test was carried out in vitro by K-B method, and adeB, deG, adej, adeR, ade S, adeE, abeM, teA and teB were collected by using K-B method. The strains of adeB, adeG, adeJ and abeM were divided into sensitive groups and drug resistant groups according to the susceptibility test, and the expression levels of ade B, adeG, adej and abeM were detected by qPCR method and compared. Results: There were 181 strains (74. 79%) and 41 strains of pus (16.94%). The first three sections were ICU (45. 04%), neurosurgery (19. 01%) and respiratory medicine (18. 18%). There were 138 males, 104 females, 157 females aged 50 to 70 years old, and 64 patients aged 50 to 70 years old; 56 (23.14%), 46 (19. 00%), 42 (17. 36%) were combined with COPD, diabetes and cardiovascular diseases. The condition of organ failure: 101 cases of respiratory failure (41. 73%), 39 cases of cardiac failure (16. 11%), 15 cases of renal failure (6. 20%), and the use of antibacterial drugs: 147 cases (60. 74%), 62 cases (25.62%) of I-linked drugs, and 33 cases (13.64%) of drug use. There were 149 cases (61. 57%) of the antibacterial drugs used during hospitalization, 87 (35. 95%) were used for carbapillenems, 86. 78% were invasive operators, 62% were artificial airway or mechanical ventilation, and the prognosis was as follows: There were 92 patients (38. 02%) who had no improvement or death during hospitalization; the average length of hospital stay was 46. 2 days. In vitro chemosensitivity test of 242 strains of Baumannii isolated from our hospital showed that the drug resistance rate was 50%, 45.04%, 29.75%, 27.27% and 3.31%, respectively. The resistance rates of ceftaxime and methamidophos were 75. 21%, 73. 55%, 71. 07%, 66. 53%, respectively. 77. 69%, 66. 12%, 65. 29%. 3. The positive rates of adeb, adeg, adej, ader, ades, adee, abem, taeta, teb, abes of 242 strains of Baumannii in our hospital were 81 82%, 85 95%, 83 47%, 81. 00%, 78. 93%, 0%, 80. 58%, 21.49%, 46. 28% and 100% respectively. The expression of adeb, adeg, adej and abem was detected by adeb, adeg, adej and abem genes, and the expression of adeb, adeg, adej and abem was tested, and the results showed that adeb, adeg, adej, abem were expressed in drug resistant group: The sensitivity groups were 21 times, 1-fold, 5-fold and 1-fold, respectively. Conclusion: 1. The distribution of the strains of Baumannii in our hospital is mainly in icu, followed by neurosurgery and respiratory medicine, mainly from sputum, the age of infected patients is large, most of them are over 70 years old, and the combination of basic diseases is more and more serious. The use of antibacterial drugs during hospitalization is much more common, and the combination phenomenon is common. In most cases of pulmonary infection, respiratory failure, invasive operation or operation may be a risk factor for the infection of Bowman. The drug resistance rate of ceftaxime, ceftripine, cefixime and drug resistance was high in the clinical isolates of Baumannii in our hospital. Carbapenem antibacterial drugs, tigecycline can be used as the first choice medicine for the clinical treatment of Baumannii infection in our hospital. The detection rate of RND (deABC, adeFGH, adeIIEF) efflux pump was generally high in Baumannii, and the detection rate of drug-resistant group was higher than that in the sensitive group. Therefore, the drug resistance of B. baumannii to antibacterial drugs could be mainly mediated by RND efflux pump, and the detection rate of adeE was 0. It is proved that there may not be adee efflux pump gene epidemic phenomenon in Baumannii in our hospital. The high expression of deABC and adeIIIa of efflux pump mediated the drug resistance of Baumannii in our hospital, while adeFGH and abeM (i.e. MATE efflux pump system) in the RAND efflux pump family mediate the further study of the drug resistance of Baumannii in our hospital.
【學(xué)位授予單位】：川北醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R446.5

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