本文選題：銅綠假單胞菌 + 耐藥基因　； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1掌握保定市銅綠假單胞菌耐藥情況及特點(diǎn),指導(dǎo)臨床醫(yī)師合理使用抗生素,延緩細(xì)菌耐藥產(chǎn)生、有效預(yù)防及控制院內(nèi)感染。2選取耐亞胺培南銅綠假單胞菌,檢測(cè)β-內(nèi)酰胺酶編碼基因種類(lèi)和外膜蛋白Opr D2基因缺失狀況,探討銅綠假單胞菌對(duì)亞胺培南藥物的耐藥機(jī)制。方法:1標(biāo)本來(lái)源收集保定市第一中心醫(yī)院住院患者2016年7月-2017年1月送檢標(biāo)本分離的銅綠假單胞菌,根據(jù)《全國(guó)臨床檢驗(yàn)操作規(guī)程》第3版的要求,用羊血培養(yǎng)基對(duì)送檢標(biāo)本進(jìn)行細(xì)菌分離和培養(yǎng),鑒定菌株采用法國(guó)梅里埃公司VITEK-2全自動(dòng)微生物鑒定儀。2藥敏實(shí)驗(yàn)將銅綠假單胞菌純化后進(jìn)行耐藥性檢測(cè),采用VITEK-2全自動(dòng)微生物鑒定儀和K-B法測(cè)定其對(duì)氨芐西林、亞胺培南等23種常用抗菌藥物的敏感性,挑選出耐亞胺培南的銅綠假單胞菌株。3耐藥基因檢測(cè)使用Maxwell?16 Cell LEV DNA Purification Kit試劑盒提取DNA,用聚合酶鏈反應(yīng)(PCR)法擴(kuò)增耐亞胺培南菌株β-內(nèi)酰胺酶基因和外膜孔道蛋白D2基因。產(chǎn)物經(jīng)2.0%瓊脂糖凝膠電泳,凝膠成像儀觀察電泳結(jié)果,與標(biāo)準(zhǔn)量DNAMarker相比較,對(duì)PCR產(chǎn)物的電泳結(jié)果進(jìn)行分析和攝像,出現(xiàn)與目的基因片段分子相當(dāng)?shù)臈l帶判為陽(yáng)性,照相保存影像。結(jié)果:1銅綠假單胞菌對(duì)常用抗菌藥物的耐藥情況測(cè)定了2016年7月到2017年1月臨床分離的122株銅綠假單胞菌對(duì)23種抗生素的耐藥性,耐藥率結(jié)果為:氨芐西林98.36%、阿米卡星10.66%、環(huán)丙沙星25.41%、慶大霉素16.39%、妥布霉素15.57%、復(fù)方新諾明96.72%、氨芐西林/舒巴坦96.72%、頭孢他啶17.21%、亞胺培南40.98%、哌拉西林18.03%、頭孢唑林97.54%、頭孢吡肟20.49%、左氧氟沙星22.95%、美羅培南23.77%、頭孢替坦95.9%、哌拉西林+他唑巴坦(常規(guī)用量)13.11%、頭孢曲松98.36%、頭孢哌酮36.07%、頭孢哌酮/舒巴坦15.57%、頭孢呋辛酯95.9%、頭孢呋辛鈉95.9%、比阿培南23.77%,呋喃妥因只用于尿液標(biāo)本的藥敏試驗(yàn),耐藥率100%。2β-內(nèi)酰胺酶編碼基因檢出情況從50株耐亞胺培南的銅綠假單胞菌檢出β-內(nèi)酰胺酶編碼基因TEM(3株)、PER(3株)、VEB(1株)、GES(2株)、OXA-2(2株)、OXA-10(3株)、CTX-M-1(3株)。3金屬β-內(nèi)酰胺酶基因檢出情況從50株耐亞胺培南的銅綠假單胞菌檢出金屬β-內(nèi)酰胺酶基因VIM(9株)、IMP(11株)、SIM(0株)、SPM(5株)、GIM(0株)。4膜孔蛋白Opr D2基因的缺失情況從50株耐亞胺培南的銅綠假單胞菌檢測(cè)出孔蛋白Opr D2基因缺失31株,缺失率為62%。結(jié)論:1保定市銅綠假單胞菌對(duì)頭孢呋辛酯、頭孢呋辛鈉、頭孢替坦、頭孢曲松、氨芐西林、復(fù)方新諾明等抗生素有很高的耐藥率。2銅綠假單胞菌對(duì)亞胺培南的耐藥率40.98%。3 50株耐亞胺培南的銅綠假單胞菌β-內(nèi)酰胺酶編碼基因檢出率為58%(29株),其中有10株同時(shí)含有兩種或兩種以上基因;Opr D2基因缺失率高(62%)。提示銅綠假單胞菌對(duì)β-內(nèi)酰胺類(lèi)抗生素的耐藥是由多種耐藥基因共同控制的,該菌具有復(fù)雜的耐藥機(jī)制。4應(yīng)加強(qiáng)銅綠假單胞菌耐藥性的監(jiān)測(cè)及耐藥基因的分子流行病學(xué)研究,防止耐藥菌株播散,提高臨床治愈率。
[Abstract]:Objective: 1 to master the drug resistance and characteristics of Pseudomonas aeruginosa in Baoding, to guide the clinicians to use antibiotics reasonably, to delay the production of bacterial resistance, to effectively prevent and control.2 in hospital infection, to select imipenem Pseudomonas aeruginosa, to detect the type of beta lactamase coding gene and the absence of Opr D2 gene in the outer membrane protein, and to explore the Pseudomonas aeruginosa. The mechanism of drug resistance of monomonas to imipenem. Methods: 1 specimen sources were collected to collect Pseudomonas aeruginosa isolated from the hospitalized patients in Baoding First Central Hospital in January -2017 July 2016. According to the requirements of the third edition of the national clinical inspection procedure, the samples were isolated and cultured with the sheep blood culture medium to identify the bacteria. The strain of Pseudomonas aeruginosa was purified by the.2 drug sensitivity test of VITEK-2 automatic microorganism identification instrument in France mereer company. The sensitivity of 23 kinds of commonly used antibiotics, such as ampicillin and imipenem, was determined by VITEK-2 automatic microorganism identification instrument and K-B method, and the Pseudomonas aeruginosa was selected. The strain.3 resistance gene was detected by using Maxwell? 16 Cell LEV DNA Purification Kit kit to extract DNA, and polymerase chain reaction (PCR) method was used to amplify the gene of beta lactamase and outer membrane D2 gene of imipenem resistant strain. The product was observed by 2% agarose gel electrophoresis and gel imaging instrument, compared with the standard quantity DNAMarker. The electrophoretic results of PCR products were analyzed and photographed, and the bands of the target genes were found to be positive, and the images were preserved. Results: 1 the resistance of Pseudomonas aeruginosa to commonly used antibiotics was tested for the resistance of 122 strains of Pseudomonas aeruginosa to 23 antibiotics from July 2016 to January 2017, and the resistance of Pseudomonas aeruginosa to January 2017 was tested. The drug rate was ampicillin 98.36%, Amikacin 10.66%, ciprofloxacin 25.41%, gentamicin 16.39%, tobramycin 15.57%, compound novamethoxine 96.72%, ampicillin / sulbactam 96.72%, ceftazidime 17.21%, imipenem 40.98%, piperacillin 18.03%, cefazolin 97.54%, cefpyoxime 20.49%, levofloxacin 22.95%, meropenem 23.77%, Ceftaetan 95.9%, piperacillin + tazobactam (conventional dosage) 13.11%, ceftriaxone 98.36%, cefoperazone 36.07%, Cefoperazone / sulbactam 15.57%, cefuroxime 95.9%, cefuroxime sodium 95.9%, amperan 23.77%, furadetin only used in urine samples, drug sensitivity test, the resistance rate 100%.2 beta lactamase gene detection from 5 0 strains of Pseudomonas aeruginosa detected beta lactamase encoding gene TEM (3 strains), PER (3 strain), VEB (1 strain), GES (2 strain), OXA-2 (2 strain), OXA-10 (3 strain), CTX-M-1 (3 strain) gene detection of metallo beta lactamase gene from 50 imipenem resistant Pseudomonas aeruginosa gene VIM (9 strain), IMP (11 strain), SIM (0) strain) SPM (5 strains), GIM (0 strains).4 membrane pore protein Opr D2 gene deletion from 50 strains of imipenem resistant Pseudomonas aeruginosa to detect the loss of pore protein Opr D2 gene deletion, the loss rate is 62%. conclusion: 1 Baoding Pseudomonas aeruginosa, cefuroxime sodium, cefuroxime, ceftriaxone, ampicillin, ampicillin, compound new nooxin and other antibiotics The resistance rate of Pseudomonas aeruginosa to imipenem.2 was highly resistant to imipenem 40.98%.3 50 strains of Pseudomonas aeruginosa resistant to Pseudomonas aeruginosa was 58% (29 strains), of which 10 had two or more than two genes, and the Opr D2 gene deletion rate was high (62%). The resistance of raw materials is controlled by multiple resistance genes, which has a complex resistance mechanism.4 should strengthen the monitoring of the resistance of Pseudomonas aeruginosa and the molecular epidemiology of resistance genes to prevent the spread of drug-resistant strains and improve the clinical cure rate.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R440

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