發(fā)布時(shí)間：2018-06-16 08:22

  本文選題：腸桿菌科細(xì)菌 + β-內(nèi)酰胺酶　； 參考：《天津醫(yī)科大學(xué)》2006年碩士論文

【摘要】：[目的]研究腸桿菌科細(xì)菌中產(chǎn)超廣譜β-內(nèi)酰胺酶(ESBLs)及質(zhì)粒介導(dǎo)AmpC酶菌株的分布。[方法]收集天津胸科醫(yī)院2004年臨床分離腸桿菌科細(xì)菌396株,以K-B法測(cè)定細(xì)菌對(duì)抗生素的敏感性;以NCCLS推薦紙片確證試驗(yàn)檢測(cè)其產(chǎn)ESBLs情況,以頭孢西丁試驗(yàn)粗篩產(chǎn)AmpC酶菌。以細(xì)菌接合試驗(yàn)輔助耐藥基因的定位。提取質(zhì)粒DNA作模板,采用聚合酶鏈反應(yīng)(PCR)和多重PCR方法及序列分析確定ESBLs及質(zhì)粒介導(dǎo)AmpC酶的基因型[結(jié)果]396株腸桿菌科細(xì)菌對(duì)氨芐西林耐藥率高達(dá)91.4%,一、二代頭孢菌素耐藥率高于40%,三代頭孢菌素中頭孢噻肟耐藥率達(dá)25.70%,慶大霉素耐藥率達(dá)26.5%,環(huán)丙沙星耐藥率達(dá)23.2%,復(fù)方磺胺耐藥率達(dá)31.8%,三耐以上的多重耐藥菌株達(dá)71.2%。其中產(chǎn)ESBLs菌株檢出率為25.0%(99/396),疑似產(chǎn)AmpC酶菌株檢出率為16.9%(67/396);產(chǎn)ESBLs菌株除對(duì)亞胺陪南仍保持較好的敏感性(99.0%)以外,對(duì)其他抗生素耐藥率均高于40%,青霉素類和一、二代頭孢菌素耐藥率均為100%,三代頭孢菌素中頭孢噻肟耐藥性最高,達(dá)79.6%。產(chǎn)ESBLs菌株接合試驗(yàn)陽(yáng)性率為40.4%(40/99),疑似產(chǎn)AmpC酶菌株接合試驗(yàn)陽(yáng)性率為10.4%(7/67),接合子藥敏試驗(yàn)證實(shí)供體菌的耐藥標(biāo)妼部分或全部傳遞給受體菌;以40株接合試驗(yàn)陽(yáng)性的產(chǎn)ESBLs菌株的質(zhì)粒DNA為模板用PCR方法進(jìn)行基因分型,結(jié)果顯示TEM、SHV、CTX-M陽(yáng)性率分別為50.0%、20.0%和67.5%,產(chǎn)2種型別ESBLs的菌株達(dá)61.3%;對(duì)7株接合試驗(yàn)陽(yáng)性疑似產(chǎn)AmpC酶菌株提取其質(zhì)粒DNA為模板,用PCR方法進(jìn)行基因分型,結(jié)果有4株為DHA型陽(yáng)性,其中2株菌同時(shí)產(chǎn)ESBLs,高度疑似為產(chǎn)超超廣譜β-內(nèi)酰胺酶(SSBL)菌株。隨機(jī)抽取5株ESBLs擴(kuò)增陽(yáng)性的PCR產(chǎn)物進(jìn)行序列分析,其結(jié)果證實(shí)2株TEM型擴(kuò)增產(chǎn)物均屬TEM-1型,經(jīng)NCBI基因庫(kù)查詢和BLASTn比對(duì),與奇異變形桿菌(AY729027)注冊(cè)的相應(yīng)序列的核
[Abstract]:[Objective] to study the distribution of ultra broad-spectrum beta lactamase (ESBLs) and plasmid mediated AmpC strains in Enterobacteriaceae bacteria. [Methods] 396 strains of Enterobacteriaceae isolated from Tianjin Chest Hospital in 2004 were collected and the sensitivity of bacteria to antibiotics was measured by K-B method; ESBLs was detected by NCCLS paper confirmation test. The bacterial conjugation test assisted the localization of the AmpC enzyme. The plasmid DNA was extracted as a template. The genotype of ESBLs and plasmid mediated AmpC was determined by the polymerase chain reaction (PCR) and multiple PCR methods. [results of the antibiotic resistance rate of ampicillin to 91.4%, one, two generation cephalosporins in]396 strains of Enterobacteriaceae. The rate of drug resistance was higher than 40%, the drug resistance rate of cefotaxime in the three generation cephalosporins was 25.70%, the drug resistance rate of gentamicin was 26.5%, the rate of ciprofloxacin resistance was 23.2%, the drug resistance rate of compound sulfonamides was 31.8%, and three resistant multiple resistant strains were 71.2%., the detection rate of ESBLs producing strains was 25% (99/396), and the detection rate of suspected producing AmpC enzyme strains was 16.9% (67/396). The resistance rate of ESBLs producing strain was higher than 40%, the resistance rate of penicillins and two generation cephalosporins was 100%, and cefotaxime in the three generation cephalosporins was the highest, and the positive rate of the 79.6%. producing ESBLs strain was 40.4% (40/99), and the suspected production of AmpC enzyme bacteria was suspected. The positive rate of the strain joint test was 10.4% (7/67). The drug sensitivity test of the zygote confirmed that the drug resistance of the donor bacteria was partly or all passed to the receptor bacteria, and the plasmid DNA of the ESBLs producing strain of 40 strains of conjugal test was genotyping by PCR method. The results showed that the positive rates of TEM, SHV, and CTX-M were 50%, 20% and 67.5%, respectively, and produced 2 types of ESBL. The strain of S was 61.3%, and the plasmid DNA was extracted from 7 strains of conjugal test, and its plasmid DNA was extracted as a template and PCR was used to genotyping. The results showed that 4 strains were DHA positive, of which 2 strains produced ESBLs at the same time and highly suspected to produce super broad-spectrum beta lactamase (SSBL) strain. 5 ESBLs amplification positive PCR products were randomly selected for sequence. The results showed that the 2 TEM type amplified products belong to the TEM-1 type, the NCBI gene pool query and the BLASTn comparison, and the corresponding sequences of the corresponding sequences registered with Proteus mirabilis (AY729027)
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R378.2

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 吳曉妹;腸桿菌科臨床耐藥株中ISCR1元件與耐藥基因水平傳播的關(guān)系研究[D];天津醫(yī)科大學(xué);2011年

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本文編號(hào)：2026031