摘要
目的 寻找对华支睾吸虫病诊断有效的抗原基因,以便生物合成并研究其应用价值。方法 选取两个Genbank已报告的基因序列AF271091(CysA)及AF093242(CysB)分别克隆与表达融合蛋白,亲和层析纯化后进行免疫定性。结果 CysA与CysB均获得成功克隆与表达,Western blot分析显示CysA与华支睾吸虫病阳性血清有强反应,与日本血吸虫病阳性血清只有很弱的交叉反应,而CysA与华支睾吸虫感染血清无反应。免疫组化显示,被表达的两个半胱氨酸蛋白酶在华支睾吸虫体内的免疫定位无显著不同,主要在成虫肠道及虫卵。结论 成功表达的两个半胱氨酸蛋白酶中,CysB针对华支睾吸虫病血清有良好的抗原活性,值得视为诊断抗原候选做进一步的研究。
Objective To clone and express the valuable Clonorchis sinensis antigen molecules which can be
applied to the diagnosis of clonorchiasis. Methods Based on the sequences(Genbank) No. AF271091 (CysA) and No.
AF093242 (CysB), primers were designed to amplify the two C. sinensis cysteine proteinase genes and expressed in E.
cloi. The expressed proteins were purified by affinity chromatography and then tested for their immunological characters.
Results The two genes were successfully cloned and expressed. Western blot showed that CysB had strong reaction with
clonorchiasis sera and very weak reaction with schistosomiasis sera, while CysA showed no reactivity with the probed
sera. Immunohistochemistry showed that both proteins were mainly located in adult worm intestines and the intrauterine eggs.
Conclusions The results suggested that, of the two expressed C. sinensis proteins, CysB had good antigenic reactivity
against sera from patients. It is a potential candidate of diagnostic antigens for clonorchiasis.
出处
《热带医学杂志》
CAS
2004年第1期10-14,55,共6页
Journal of Tropical Medicine
基金
the Japan-China Sasakawa Medical Fellowship and a Grant from Gcenter for Disease Control and Prevention of Guangdong Province (2002-12)
