贝氏柯克斯体5种重组表面蛋白抗原包被酶联免疫吸附试验的特异性和敏感性分析  

Specificity and sensitivity of enzyme-linked immunosorbent assay with five recombinant surface protein antigens of Coxiella burnetii

作　　者：汪鹏程[1,2] 熊小路[1] 焦俊[1] 龚文平[1] 杨小梅 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,病原生物生物安全国家重点实验室,北京100071 [2]中国人民解放军第105医院检验科,合肥230031

出　　处：《中国人兽共患病学报》2015年第12期1111-1115,1119,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.31170161);国家科技重大项目(No.2013ZX10004803)联合资助~~

摘　　要：目的使用贝氏柯克斯体5种重组表面蛋白作为包被抗原的酶联免疫吸附试验用于血清学检测的特异性和敏感性分析。方法本研究建立由贝氏柯克斯体的Com1、GroEL、Mip、OmpH、RplL重组表面蛋白抗原分别包被的酶联免疫吸附试验(ELISA)方法,然后分别检测贝氏柯克斯体实验感染小鼠血清和Q热患者血清中IgG特异性抗体。结果 8份贝氏柯克斯体感染小鼠血清中除1份血清与RplL反应为阴性外其它均为阳性;11份Q热患者血清中除1份血清与RplL、1份血清与OmpH反应为阴性外,其它血清反应均为阳性。将这5种重组蛋白分别与莫氏立克次体、黑龙江立克次体、恙虫病东方体实验感染小鼠血清反应,除1份黑龙江立克次体感染小鼠血清与GroEL反应为阳性外,其它血清反应均为阴性。另外,Com1对斑疹伤寒、斑点热、恙虫病患者血清的阳性反应率平均为22.2%,GroEL为25.0%,Mip为25.0%,OmpH为25.0%,RplL为13.9%。结论这些结果证明这5种重组蛋白抗原均可被贝氏柯克斯体感染血清所识别。所建立的ELISA方法具有良好特异性和敏感性,可用作Q热血清学诊断。In order to evaluate the usefulness of recombinant proteins of Coxiella burnetti as serodiagnostic markers for Q fever,the enzyme-linked immunosorbent assay（ELISA）coated with each of five recombinant surface protein antigens（Com1,GroEL,Mip,OmpH,or RplL）of Coxiella burnetii was assessed for specificity and sensitivity in detection of specific IgG antibodies in sera from mice experimentally infected with C.burnetii and patients suffered from Q fever.As a result,all of 8sera fromC.burnetii-infected mice positively reacted with the five recombinant proteins,except for one serum which negatively reacted with RplL,while all of 8sera from mice infected with Rickettsia mooseri,Rickettsia heilongjiangensis,or Orientia tsutsugamshi negatively reacted with the five proteins,except for one serum which positively reacted with GroEL.In addition,the average positive ratio of 11 sera from patients with endemic typhus,13 sera from patients with spotted fever,and 12 sera from patients with tsutsugamushi disease were 22.2% for reaction to Com1,25.0% for GroEL,25.0% for Mip,25.0% for OmpH,and 13.9%for RplL,respectively.Results suggest that the ELISA with Com1,GroEL,Mip,OmpH,or RplL has high specificity and sensitivity for serological diagnosis of Q fever.

关 键 词：贝氏柯克斯体 Q热 表面蛋白 酶联免疫吸附试验 

分 类 号：R446.5[医药卫生—诊断学]