摘要
目的探讨细粒棘球蚴重组抗原(rEgmMDH)对囊型包虫病的血清学诊断价值。方法将已构建的原核表达菌株mMDH/pET-28a/BL21■lysS,用IPTG诱导表达,经亲和层析纯化获得高纯度rEgmMDH,以rEgmMDH为抗原,应用ELISA和Western-blotting方法对40例手术确诊的囊型包虫病病人血清进行了回顾性检测与分析。结果 ELISA和Western-blotting方法检测囊型包虫病病人血清阳性率为92.5%(37/40);Western-blotting方法显示囊型包虫病病人血清可特异性识别重组抗原,泡球蚴病人汇集血清不能识别重组抗原。结论 rEgmMDH具有较好的抗原性,可作为囊型包虫病血清学检测的候选诊断抗原。
Objective To explore the sensibility and the antigenic specificity of Echinococcus granulosus recombinant antigen rEgmMDH through ELISA and Western-blotting and to evaluate its useess in serodiagnosis of human cystic echinococcosis.Methods A previously constructed rEgmMDH prokaryotic expression plasmid vector（pET28a/rEg mMDH） was transformed into E.coliBL21（DE3）.Protein expression was induced with IPTG and the expressed rEg mMDH was purified using His-tagged affinity chromatography column.40 serum samples from surgically confirmed patients with cystic echinococcosis（CE） were analyzed by ELISA and Western-blotting using rEg mMDH as diagnostic antigen.At the same time,the AE patients pooled serum were collected to recognize the rEgmMDH by Western-blotting method.Results 37（92.5 %） of 40 serum samples were positive both in rEg mMDH ELISA and rEg mMDH Western-blotting and the remaining 3 sero-negative serum samples from CE patients with single cyst in liver were diagnosed for the first time.The AE patients pooled serum could not recognize the rEgmMDH specially while the CE patients pooled serum could.Conclution The study shows that rEg mMDH is highly useful for serodiagnosis of human cystic echinococcosis while possessing high antigenic sensibility and specificity can act as a promising molecular diagnostic agent for CE.
出处
《宁夏医科大学学报》
2011年第3期216-218,共3页
Journal of Ningxia Medical University
基金
宁夏医科大学校级课题
