金黄色葡萄球菌EsxB蛋白的表达、纯化及应用  

Expression, purification and application of EsxB protein in Staphylococcus aureus

作　　者：杜鸿[1] 张萍[1] 沈海英[1] 王敏[1] 戴晓莉[1] 

机构地区：[1]苏州大学附属第二医院检验科,215004

出　　处：《中华预防医学杂志》2012年第4期364-366,共3页Chinese Journal of Preventive Medicine

基　　金：江苏省卫生厅科研项目（H201014）

摘　　要：目的建立EsxB蛋白的表达、纯化方法，了解金黄色葡萄球菌（简称金葡菌）感染者血清EsxB抗体阳性比例及其耐药特征。方法同源重组构建EsxB蛋白原核表达体系，诱导表达后果用超小波法破碎细菌，以镍离子螯合层析柱纯化EsxB蛋白；以EsxB为抗原，应用间接ELISA法检测78例临床金葡菌感染患者血清EsxB抗体；应用全自动细菌鉴定分析仪检测相应的78株金葡菌分离菌株的耐药情况。结果成功构建EsxB原核表达体系并纯化得剑EsxB蛋白；28．2％（22／78）的金葡菌感染患者血清EsxB抗体阳性，来自EsxB抗体阳性患着金葡菌分离株，其多重耐约菌株的的比例（100．0％，22／22）和耐甲氧两林菌株的比例（77．3％，17／22）均高于来自EsxB机体阴性患者的金葡菌分离株洲[分别为35．7％（20／56）和21．4％（12／56）]（P值均〈0．01）结论建立了EsxB蛋白的表达、纯化疗法；分离自EsxB抗体阳性者的金葡菌均为耐药菌株，且大多数对甲氧西林耐药。Objective This study aimed to establish the method of expression and purification of EsxB protein, explore the EsxB antibody-positive Staphylococcus aureus （ S. aureus） clinical infection status and relevance of drug resistanee. Methods Construeted EsxB prokaryotic expression system by homologous recombination, Ni2＋ column was used to purify EsxB protein; and then ELISA was used to detect the anti- EsxB antihodies in serum of 78 patients with S. aureus infection; antimicrobial susceptibility of related S. aureus strains by automatic bacterial identification analyzer. Results EsxB prokaryotic protein expression svstem was constructed and EsxB protein was purified successfully; anti-EsxB antibodies were present in the serum of patients with S. aureus infection up to 28.21% （22/78）. The proportion of multi-drag resistant and methieillin-resistant S. aureus strains isolated from anti-EsxB antibodies positive patients were 100.0% （22/22）, 77.3% （17/22） , respectively, which were statistically higher than those strains isolated from anti-EsxB antibody-negative patients （ 35.7% （ 20/56 ） and 21.4% （ 12/56 ）, respectively ） （ all P values 〈 0. 01 ）. Conclusion Method for expression and purification of EsxB protein was established. All the S. aureus strains isolated from EsxB antibody- positive patients were multidrug resistant strains and most of them were resistant to methicillin.

关 键 词：葡萄球菌 金黄色 多重耐药 蛋白纯化 EsxB抗体 

分 类 号：R378[医药卫生—病原生物学]