太原地区产超广谱β-内酰胺酶大肠埃希菌的流行趋势和耐药机制
发布时间：2018-11-25 08:51
【摘要】： 目的 分析太原地区产ESBL大肠埃希菌流行趋势和耐药机制,为指导临床合理使用抗生素,减少耐药菌株在本地区扩散及流行病研究提供实验依据和指导。 方法 1对临床分离的耐药菌株进行纸片扩散法ESBL表型筛选,将筛选出68株ESBL表型阳性菌重新进行生化鉴定、药敏实验。 2用PCR方法对产ESBL大肠埃希菌进行分型并测序。 3用三维实验、多重PCR的方法对耐头孢西丁的产ESBL大肠埃希菌进行AmpCβ-内酰胺酶的检测。 4 SDS-PAGE对部分菌株外膜蛋白进行分离鉴定。 5 ERIC—PCR的方法流行病学分型。 6 PCR方法对整合酶及其基因盒进行分析并测序。 结果 1临床收集耐药大肠埃希氏菌株中,通过ESBL表型确证试验共筛选出68株ESBL阳性菌株。其中门诊病人10株,住院病人58株。其中外科病房分离占2/3。 2 PCR结果TEM型67株为阳性,SHV型04LE030阳性,测序为SHV12, CTX-M2、OXA1、OXA2、OXA10引物PCR电泳未见阳性。CTX-M1阳性21株,CTX-M9阳性48株。 3 AMPC三维实验04LE030弱阳性,多重PCR结果04LE030阳性带在400BP左右,测序为DHA1。 4 SDS-PAGE可见04LE048出现OmpF缺失出现大分子蛋白条带。 5.ERIC—PCR的方法显示不同医院菌株流行病学分型有差异。 6整合酶ⅠPCR阳性39株,整合酶ⅡPCR阳性1株,其基因盒有两种结构,均包括 氨基糖苷类抗生素和磺胺类抗生素的耐药的基因。结论 1产ESBL大肠埃希菌引起的感染以泌尿系统和呼吸道感染为主,住院病人以外科病房为主,门诊病人泌尿系统老人多见。 2本地区产ESBL大肠埃希菌以CTX-M9群、CTX-M1群为主。表型筛选时可使用头孢噻肟和氨曲南,确证可只使用头孢噻肟和头孢噻肟+棒酸。单头孢他定和头孢他定+棒酸表型确证试验阳性的大肠高度怀疑为SHV型。 3本地区同时产ESBL和AmpC大肠埃希菌还不多见;头孢西丁完全耐药(直径=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，
本文编号：2355513
[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，
本文编号：2355513
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