本文選題：大腸埃希菌 + 代謝組學(xué)��； 參考：《川北醫(yī)學(xué)院》2015年碩士論文

【摘要】：目的:統(tǒng)計(jì)分析大腸埃希菌(Escherichia coli,E.coli)感染者臨床資料及該菌株對(duì)三類15種抗菌藥物的耐藥性;用氣相色譜-質(zhì)譜聯(lián)用(GC-MS)代謝組學(xué)的方法尋找與耐藥相關(guān)的差異性代謝產(chǎn)物和代謝網(wǎng)絡(luò),并進(jìn)一步探討其與耐藥之間的關(guān)系。方法:收集附屬醫(yī)院微生物室2012年12月至2014年10月臨床非同一患者同一部位重復(fù)分離的E.coli共126株,采用瓊脂二倍稀釋法檢測15種臨床常用抗菌藥物的最低抑菌濃度(minimal inhibitory concentration,MIC),并統(tǒng)計(jì)分析其感染者臨床資料和對(duì)抗菌藥物的耐藥性;篩選出對(duì)上述抗菌藥物全部敏感的6株(S組)和耐藥譜一致的4株(R組)作為代謝組學(xué)研究對(duì)象,利用GC-MS代謝組學(xué)的方法分析S組和R組中E.coli的代謝特征譜的差異,結(jié)合相關(guān)代謝通路數(shù)據(jù)庫和藥物機(jī)制尋找與耐藥相關(guān)的胞內(nèi)差異性代謝產(chǎn)物和代謝網(wǎng)絡(luò)。結(jié)果:126例患者病史資料提示平均年齡為31.57±8.26歲,女性占51.59%(65/126)。病人有基礎(chǔ)性疾病者占10.32%(13/126),手術(shù)治療患者占23.89%(30/126)。入院前或入院后使用2種或以上的抗菌藥物治療者占56.35%(71/126)。平均住院時(shí)間為13.6天。標(biāo)本來源科室分布:泌尿科(泌尿外科和腎內(nèi)科)占42.86%(54/126),普外科和肛腸科占28.57%(36/126),婦產(chǎn)科占14.29%(18/126),呼吸科占8.73%(11/126)。標(biāo)本取材來源以尿液和糞便最多,占54.76%(69/126),其次是手術(shù)創(chuàng)面和泌尿生殖道分泌物26.19%(33/126),痰液11.11%(14/126)。126株E.coli對(duì)頭孢唑林耐藥率最高(71.43%),頭孢呋辛、慶大霉素的耐藥率均在60%以上,對(duì)亞胺培南/西司他丁100%敏感,頭孢米諾、頭孢西丁和阿米卡星敏感性均在90%以上,敏感菌株占17.46%(22/126),多重耐藥(MDR)菌株占74.6%(94/126)。磷酸、丁二酸、磷酸乙醇胺、甘油、順-10-十七碳烯酸是S組和R組的差異性代謝產(chǎn)物,在R組的含量均低于S組。它們參與三羧酸循環(huán)、氧化磷酸化、氨基酸代謝、磷脂代謝等代謝通路。結(jié)論:(1)我院臨床分離E.coli對(duì)所試三類15種臨床常用抗菌藥物敏感率為17.46%,對(duì)兩類抗菌藥物耐藥率為9.74%,多重耐藥率為74.6%;亞胺培南、頭孢米諾、頭孢西丁和阿米卡星是治療耐藥E.coli感染的理想選擇,尤以亞胺培南為最。(2)磷酸、丁二酸、磷酸乙醇胺、甘油、順-10-十七碳烯酸是S組和R組的差異性代謝產(chǎn)物,它們在R組的含量皆低于S組,可能與R組中細(xì)菌在抗菌藥物長期壓力下的低代謝水平有關(guān)。(3)R組中丁二酸含量低可能是E.coli中TCA代謝失衡所致,進(jìn)而干擾細(xì)胞的能量代謝,這大概與細(xì)菌的自我保護(hù)機(jī)制有關(guān)。(4)R組中磷酸、磷酸乙醇胺、甘油含量低可能是E.coli為逃避抗菌藥物打擊致使它們參與了細(xì)胞膜的空間構(gòu)象改變,為其耐藥性的產(chǎn)生提供必要的生物學(xué)基礎(chǔ)。
[Abstract]:Objective: to analyze the clinical data of Escherichia coli (E.coli) infection and the resistance of the strain to three kinds of 15 kinds of antibiotics; to find the differential metabolites and metabolic networks related to drug resistance by gas chromatography-mass spectrometry (GC-MS), and to further explore the relationship between them and drug resistance. Methods: a total of 126 E.coli isolates from the same site in the hospital from December 2012 to October 2014 were collected, and the minimum inhibitory concentration (minimal inhibitory concentration, MIC) of 15 kinds of commonly used antibiotics was detected by agar two times dilution method, and the clinical data and antagonistic bacteria of the infected persons were analyzed and analyzed. Drug resistance; screening 6 strains (group S) and 4 strains (group R) that are all sensitive to the above antibiotics (group S) and resistance spectrum (group R). The metabolic characteristics of E.coli in group S and R group were analyzed by GC-MS metabolomics, and the related metabolic pathway database and drug mechanism were used to find the cell related to drug resistance. Results: the internal differential metabolites and metabolic networks. Results: the average age of 126 patients was 31.57 + 8.26 years, 51.59% (65/126), 10.32% (13/126) and 23.89% (30/126). 2 or more antibiotics were used before or after admission (71/126). The duration of hospitalization was 13.6 days. The distribution of specimen sources: Urology (Department of Urology and Nephrology) accounted for 42.86% (54/126), Department of general surgery and anorectal Department accounted for 28.57% (36/126), gynecologic and obstetrics accounted for 14.29% (18/126), 8.73% (11/126). The specimens were collected from urine and feces, accounting for 54.76% (69/126), followed by surgical wound and genitourinary tract. Secretion 26.19% (33/126), sputum 11.11% (14/126).126 strain E.coli was the highest (71.43%) resistant to cefazolin, and cefuroxime, gentamicin resistance rates were above 60%, imipenem / cefastatin 100%, ceftriaxone, cefoxitin and Amikacin were more sensitive than 90%, sensitive strains accounted for 17.46% (22/126), multidrug resistance (MDR). The strains accounted for 74.6% (94/126). Phosphoric acid, succinic acid, ethanolamine phosphate, glycerol, and CIS -10- seventeen carboxenoic acid were different metabolites in group S and R, and in R group were lower than that in group S. They were involved in the metabolic pathways of three carboxylic acid cycle, oxidative phosphorylation, amino acid metabolism, phospholipid metabolism, etc. conclusion: (1) the clinical separation of E.coli in our hospital on the three class 15 faces of the trial. The susceptibility rate of common antibiotics in bed was 17.46%, the rate of resistance to two types of antibiotics was 9.74%, and the rate of multidrug resistance was 74.6%. Imipenem, cephalosporin, cefoxitin and Amikacin were the ideal choice for the treatment of drug-resistant E.coli infection, especially imipenem. (2) phosphoric acid, succinic acid, ethanolamine phosphate, glycerol, and CIS -10- seventeen carbonic acid were the S group and the S group. The difference of metabolites in group R, which are lower in group R than in group S, may be related to the low metabolism level of bacteria in group R under long-term pressure of antimicrobial agents. (3) low content of succinic acid in group R may be caused by metabolic imbalance of TCA in E.coli, thereby interfering with cell energy metabolism, which is probably related to the self-protection mechanism of bacteria. (4) R group. The low content of phosphoric acid, ethanolamine phosphate and glycerin may be that E.coli is an escape from the anti antibacterial drug attack that causes them to participate in the spatial conformation change of the cell membrane, and provides the necessary biological basis for the production of its resistance.
【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】：2015
【分類號(hào)】：R446.5

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