贝氏柯克斯体血清学反应蛋白芯片的研制  

Protein Microarray of Seroreactive Proteins of Coxiella burnetii

作　　者：熊小路[1] 王锡乐[1] 孟艳芬[1] 齐永[1] 温博海[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《寄生虫与医学昆虫学报》2011年第2期90-94,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家科技重大专项（重大传染病预防与控制课题2008ZXl0004-002）;国家自然科学青年基金项目（No.30901369）

摘　　要：贝氏柯克斯体为Q热的病原菌。本研究对19个贝氏柯克斯体血清学反应阳性蛋白基因进行PCR扩增，将扩得的基因片段与表达载体连接，将重组质粒转化大肠杆菌并表达目的蛋白，采用Ni-NTA亲和层析柱纯化目的蛋白。将纯化的重组蛋白点在醛基化玻片上制备蛋白芯片。将贝氏柯克斯体新桥株实验感染小鼠血清分别对蛋白芯片进行血清学分析，结果显示能与贝氏柯克斯体感染1、2、3、4周小鼠血清反应的蛋白数分别是0、16、19和18个。结果说明该芯片上的重组蛋白具有贝氏柯克斯体感染血清反应特异性，该芯片分析有望用于Q热的血清学诊断。Coxiella burnetii is the etiological agent of Q fever. Nineteen genes encoding sero-reactive proteins amplified by PCR were inserted into vectors and expressed in E. coll. The recombinant proteins were purified by Ni-NTA affinity chromatography and used to prepare protein microarray. The protein microarray was probed with sera from mice experimentally infected with C. burnetii, and 0, 16, 19 and 18 recombinant proteins on the microarray were recognized by mouse sera collected on week 1, 2, 3 and 4 post-infection, respectively. The results indicated that the proteins on the microarray are specifically reactive to antibodies against C. burnetii, and may be used for Q fever sero-diagnosis.

关 键 词：贝氏柯克斯体 Q热 血清反应阳性蛋白 蛋白芯片 

分 类 号：Q51[生物学—生物化学]