發(fā)布時(shí)間：2018-11-25 08:51

【摘要】： 目的 分析太原地區(qū)產(chǎn)ESBL大腸埃希菌流行趨勢(shì)和耐藥機(jī)制,為指導(dǎo)臨床合理使用抗生素,減少耐藥菌株在本地區(qū)擴(kuò)散及流行病研究提供實(shí)驗(yàn)依據(jù)和指導(dǎo)。 方法 1對(duì)臨床分離的耐藥菌株進(jìn)行紙片擴(kuò)散法ESBL表型篩選,將篩選出68株ESBL表型陽(yáng)性菌重新進(jìn)行生化鑒定、藥敏實(shí)驗(yàn)。 2用PCR方法對(duì)產(chǎn)ESBL大腸埃希菌進(jìn)行分型并測(cè)序。 3用三維實(shí)驗(yàn)、多重PCR的方法對(duì)耐頭孢西丁的產(chǎn)ESBL大腸埃希菌進(jìn)行AmpCβ-內(nèi)酰胺酶的檢測(cè)。 4 SDS-PAGE對(duì)部分菌株外膜蛋白進(jìn)行分離鑒定。 5 ERIC—PCR的方法流行病學(xué)分型。 6 PCR方法對(duì)整合酶及其基因盒進(jìn)行分析并測(cè)序。 結(jié)果 1臨床收集耐藥大腸埃希氏菌株中,通過ESBL表型確證試驗(yàn)共篩選出68株ESBL陽(yáng)性菌株。其中門診病人10株,住院病人58株。其中外科病房分離占2/3。 2 PCR結(jié)果TEM型67株為陽(yáng)性,SHV型04LE030陽(yáng)性,測(cè)序?yàn)镾HV12, CTX-M2、OXA1、OXA2、OXA10引物PCR電泳未見陽(yáng)性。CTX-M1陽(yáng)性21株,CTX-M9陽(yáng)性48株。 3 AMPC三維實(shí)驗(yàn)04LE030弱陽(yáng)性,多重PCR結(jié)果04LE030陽(yáng)性帶在400BP左右,測(cè)序?yàn)镈HA1。 4 SDS-PAGE可見04LE048出現(xiàn)OmpF缺失出現(xiàn)大分子蛋白條帶。 5.ERIC—PCR的方法顯示不同醫(yī)院菌株流行病學(xué)分型有差異。 6整合酶ⅠPCR陽(yáng)性39株,整合酶ⅡPCR陽(yáng)性1株,其基因盒有兩種結(jié)構(gòu),均包括 氨基糖苷類抗生素和磺胺類抗生素的耐藥的基因。結(jié)論 1產(chǎn)ESBL大腸埃希菌引起的感染以泌尿系統(tǒng)和呼吸道感染為主,住院病人以外科病房為主,門診病人泌尿系統(tǒng)老人多見。 2本地區(qū)產(chǎn)ESBL大腸埃希菌以CTX-M9群、CTX-M1群為主。表型篩選時(shí)可使用頭孢噻肟和氨曲南,確證可只使用頭孢噻肟和頭孢噻肟+棒酸。單頭孢他定和頭孢他定+棒酸表型確證試驗(yàn)陽(yáng)性的大腸高度懷疑為SHV型。 3本地區(qū)同時(shí)產(chǎn)ESBL和AmpC大腸埃希菌還不多見;頭孢西丁完全耐藥(直徑=6mm)
[Abstract]:Objective to analyze the epidemic trend and drug resistance mechanism of ESBL producing Escherichia coli in Taiyuan, so as to provide experimental basis and guidance for clinical rational use of antibiotics and reducing the spread of resistant strains in this area. Methods 1 the phenotype of ESBL was screened by disc diffusion method. 68 strains of ESBL phenotypic positive strains were screened for biochemical identification and drug sensitivity test. 2 PCR method was used to type and sequence ESBL producing Escherichia coli. 3 AmpC 尾 -lactamases were detected by using three dimensional experiment and multiple PCR method in ESBL producing Escherichia coli resistant to cefoxitin. 4. The outer membrane proteins of some strains were isolated and identified by SDS-PAGE. 5Method of ERIC-PCR epidemiological typing. The integrase and its gene cassette were analyzed by PCR and sequenced. Results (1) 68 ESBL positive strains were screened by ESBL phenotypic confirmatory test in clinical collection of drug-resistant Escherichia coli strains. There were 10 outpatients and 58 inpatients. Surgical ward separation accounted for 2 / 3 of the total. 2the results of PCR showed that 67 strains of TEM type were positive, and SHV type 04LE030 was positive. SHV12, CTX-M2,OXA1,OXA2,OXA10 primer PCR electrophoresis was not positive, CTX-M1 was positive in 21 strains and CTX-M9 was positive in 48 strains. 3 04LE030 was weakly positive in AMPC three-dimensional experiment, and 04LE030 positive band was about 400BP in multiple PCR results. The sequence was DHA1.. 4 SDS-PAGE showed that 04LE048 had OmpF deletion and macromolecular protein bands. The method of 5.ERIC-PCR showed that there were differences in epidemiological typing of strains in different hospitals. (6) 39 strains of integrase 鈪，

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