Development of an easy and quick serodiagnostic method for important parasitosis using geratine particles coated with recombinant parasite antigens

使用涂有重组寄生虫抗原的吉拉汀颗粒开发一种简单快速的重要寄生虫病血清诊断方法

• 批准号: 07557206
• 负责人: SENDO Fujiro
• 金额: $ 3.2万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557206/
• 关键词: Schistosoma japonicum; recombinant protein; geratine particle; an easy and quick serodiagnostic method; monoclonal antibody; リコンビナント抗原; シャーガス病; リーシュマニア症; 日本住血吸虫症

Schistosomiasis japonicum is one of the important infectious diseases in the world which is endemic mainly in China and Philippines. Up to now, there are some diagnostic methods such as ELISA,IHA,and IFAT etc. However, these need special reagents, expensive instruments, and skillful techniques. Therefore, they can not be used widely in the endemic fields. To resolve these problems, we tried to develop a novel, easy, and quick serodiagnostic method using by gelatin particles coated with recombinant parasite antigens.We successed to analyze a whole cDNA sequence of Schistosoma japonicum adult antigen recognized by SJA111 mAb, established in our department. The cDNA sequence was almost the same as a known one, except 4 base pairs. However, there is a no report about this recombinant protein. Therefore, we produced the recombinant proteins from BL21 (DE3) E.coli transformed with Pet23b-SJAcDNA.The recombinant proteins were purified with an affinity column, subjected to SDS 12% PAGE,and immunoblotted with SJA111 mAb. The recombinant proteins reacted with SJA111 mAb. Therefore, we produced and purified in a small scale the recombinant proteins recognized by SJA111 mAb. We are now trying to obtain large amounts of purified recombinant proteins. And then, we will check whether the gelatin particles coated with the purified recombinant proteins are useful tool or not for serodiagnosis of schistosomiasis japonicum by using patient sera of many kinds of parasitosis, including schistosomiasis, from Laos.

日本血吸虫病是世界性的重要传染病之一，主要流行于中国和菲律宾。目前的诊断方法有ELISA、IHA、IFAT等，但这些方法需要特殊的试剂、昂贵的仪器和熟练的技术。因此，它们不能广泛应用于地方病领域。为了解决这些问题，我们尝试建立了一种新的、简便、快速的血清学诊断方法-该cDNA序列与已知的cDNA序列基本相同，只有4个碱基对不同。然而，目前还没有关于该重组蛋白的报道。因此，我们用Pet 23 b-SJAcDNA转化BL 21（DE 3）大肠杆菌，获得重组蛋白，用亲和层析柱纯化，SDS12%PAGE电泳，用SJA 111单克隆抗体进行免疫印迹。重组蛋白与SJA 111单抗反应。因此，我们在小规模生产和纯化的重组蛋白识别的SJA 111单克隆抗体。我们现在正在努力获得大量纯化的重组蛋白。然后，我们将检查是否与纯化的重组蛋白包被的明胶颗粒是有用的工具或不使用多种寄生虫病，包括血吸虫病，从老挝的患者血清，日本血吸虫病的血清诊断。