日本血吸虫谷胱甘肽-S-转移酶在虫卵阶段的定位  

Schistosoma japonicum glutathione-S-transferase(Sj26GST):expression,localization and its evaluation in diagnosis for schistosomiasis

作　　者：王志成[1,2] 徐元宏[2] 汪学龙[1] 李敏[1] 罗飞[1] 郑美娟[1] 罗庆礼[1] 沈继龙[1] 

机构地区：[1]安徽医科大学病原生物学教研室,教育部重要遗传病基因资源利用重点实验室和安徽省重要遗传病基因资源利用重点实验室,合肥230032 [2]安徽医科大学第一附属医院检验科,合肥230022

出　　处：《安徽医科大学学报》2007年第1期1-4,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家高技术研究发展计划(863计划)(编号:2004AA2Z3570);国家自然科学基金(编号:30571631)

摘　　要：目的探讨谷胱甘肽-S-转移酶(Sj26GST)在虫卵阶段的表达、定位和重组蛋白(rSj26GST)的免疫诊断价值。方法从感染日本血吸虫尾蚴6周的兔肝脏中分离虫卵,分别用RT-PCR和免疫印迹(Western blotting)法检测Sj26GST基因在虫卵阶段的转录和翻译;同时用兔肝做免疫组化观察Sj26GST在虫卵内的分布。利用纯化的rSj26GST和日本血吸虫可溶性虫卵抗原(SEA),采用间接ELISA法检测急、慢性血吸虫病患者和正常人血清。结果RT-PCR从血吸虫虫卵中扩增670bp的基因片断,Western blotting证明在虫卵阶段Sj26GST的表达;同时免疫组化显示Sj26GST主要分布在虫卵内毛蚴两边的侧腺。rSj26GST检测急、慢性血吸虫病患者和正常人血清的阳性率分别为92%、80%和0;用SEA检测以上标本,阳性率分别为96%、84%和2%。结论Sj26GST是SEA的主要组分之一,能刺激机体产生高滴度的抗体;rSj26GST为抗原诊断日本血吸虫病显示出高度的敏感性和特异性,具有实用价值。Objective To investigate the expression and localization of glutathione-S-transferase（Sj26GST）in eggs stage of Schistosoma japonicum and its evaluation on diagnosis of shistosomiasis. Methods The eggs were separated from liver tissues of the rabbits which had been infected with S. japonicum on day 42 after infection. The transcription and translation of Sj26GST gene in eggs were confirmed by RT-PCR and Western blotting by using total RNA and solubale egg antigen（SEA）. Meanwhile, Immuno-loealization of SjGST within eggs was performed with monoelonal antibody to Sj26GST. Sera from patient with schistosomiasis were detected with purified rSj26GST and SEA by indirect ELISA. Results Consistent with the expected size, 670 bp of amplified product of RT-PCR was ob- tained and expression of Sj26GST gene in eggs was recognized by Western blotting. The Sj26GST was predominantly localized in lateral gland of miraeidia in eggs. The results of ELISA showed that the positive rates of anti- Sj26GST antibodies in sera from acute and chronic eases of schistosomiasis were 92% and 80% , while the positive rates of anti-SEA antibodies were 96% and 84% , respectively. No cross-reactivity was found between rSj26GST and normal sera, while slight nonspeeeifie reaction in sera from normal persons was noted by SEA. Conclusion The study shows that the rSj26GST has a potential as a recombinant reagent for the diagnosis of schistosomiasis.

关 键 词：血吸虫 日本 血吸虫病/诊断 寄生虫虫卵计数 

分 类 号：R345.5[医药卫生—基础医学] R383.24