rSj26GST-Sj32融合蛋白-IgG-ELISA用于慢性日本血吸虫病诊断的研究  

Diagnosis of Chronic Schistosomiasis Japonicum with the Recombinant Sj26GST-Sj32 Fusion Protein by ELISA

作　　者：蔡世飞[1] 李文桂[1] 王敏[1] 

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《四川大学学报（医学版）》2011年第3期331-334,共4页Journal of Sichuan University(Medical Sciences)

基　　金：重庆市科委地方病重大专项基金(2008AB5055;2008AB5008;2008AB5054)资助

摘　　要：目的探讨rSj26GST-Sj32-IgG-ELISA用于慢性日本血吸虫病的诊断价值。方法将含有重组质粒pET32α-Sj26GST-Sj32的大肠埃希菌BL21(DE3)用异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达,表达产物进行SDS-PAGE电泳,用Ni-NTA试剂盒纯化重组Sj26GST-Sj32(rSj26GST-Sj32)融合蛋白。常规方法制备日本血吸虫成虫抗原(SjAWA)。用rSj26GST-Sj32融合蛋白和SjAWA作为包被抗原建立ELISA检测慢性日本血吸虫病患者的血清特异性IgG抗体,同时以华支睾吸虫病、卫氏并殖吸虫病、泡型棘球蚴病、囊型棘球蚴病、乙型肝炎、肺结核患者血清和健康人血清作为对照。结果 rSj26GST-Sj32融合蛋白检测慢性日本血吸虫病患者的敏感性和特异性分别为95.00%(38/40)和97.67%(42/43),而SjAWA分别为92.50%(37/40)和97.67%(42/43)。两种抗原的检出率差异无统计学意义(P均>0.05)。两种抗原与华支睾吸虫病和卫氏并殖吸虫病患者血清均有不同程度的交叉反应。rSj26GST-Sj32融合蛋白与泡型棘球蚴病患者血清无交叉反应,但SjAWA与该病2例患者血清有交叉反应,两者比较差异无统计学意义(P>0.05)。结论 rSj26GST-Sj32-IgG-ELISA可用于慢性日本血吸虫病的免疫诊断。Objective To study the feasibility of rSj26GST-Sj32-IgG-ELISA for diagnosis of chronic schistosomiasis japonicum.Methods The Escherichia coli BL21（DE3） with recombinant plasmid pET32α-Sj26GST-Sj32 were induced with isopropy-β-D-thiogalactopyranosid（IPTG）,and the expression product was analyzed by SDS-PAGE and purified by Ni-NTA kits.Schistosoma japonicum（Sj） adult worm antigen（SjAWA） was produced by routine method.The IgG antibodies were detected with the sera of chronic schistosomiasis japonicum by ELISA using recombinant Sj26GST-Sj32（rSj26GST-Sj32） fusion protein and SjAWA,while the controls included the sera of patients with clonorchiasis,paragonimiasis westermani,alveolar echinococcosis,cystic echinococcosis,hepatitis B,pulmonary tuberculosis and healthy people.Results The sensitivity and specificity of rSj26GST-Sj32 fusion protein were 95.00%（38/40） and 97.67%（42/43） respectively,they were 92.50%（37/40） and 97.67%（42/43） respectively in SjAWA groups.There were no difference in sensitivity and specificity between rSj26GST-Sj32 and SjAWA（P〉0.05）.There were different cross reactions in clonorchiasis sinensis and paragonimiasis westermani between the two methods.The cross reaction with SjAWA was 20.00%（2/10） in patients with alveolar echinococcosis,but no cross reaction with rSj26GST-Sj32（P〉0.05）.Conclusion rSj26-Sj32-IgG-ELISA probably could be applied to immunodiagnosis for chronic schistosomiasis japonicum.

关 键 词：酶联免疫吸附试验 rSj26GST-Sj32融合蛋白 IGG 

分 类 号：R532.21[医药卫生—内科学]