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

1996 Fiscal Year Final Research Report Summary

• Project/Area Number: 07557206
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 試験
• Research Field: 寄生虫学(含医用動物学)
• Research Institution: Yamagata University
• Principal Investigator: SENDO Fujiro Yamagata University, School of Medicine, Professor, 医学部, 教授 (80091833)
• Co-Investigator(Kenkyū-buntansha): FUJINO Ryuuichi Fujirebio Inc., Central Research Laboratory, 中央研究所, 室長 ; ARAKI Yoshihiko Yamagata Univetsity, School of Medicine, Assistant Professor, 医学部, 助手 (70250933) ; WATANABE Tadashi Yamagata University, School of Medicine, Assistant Professor, 医学部, 助手 (60113990) ; YAMASHITA Takao Yamagata University, School of Medicine, Assistant Professor, 医学部, 助手 (80018928) ; SAITO Susumu Yamagata University, School of Medicine, Associate Professor, 医学部, 助教授 (50034004)
• Project Period (FY): 1995 – 1996
• Keywords: Schistosoma japonicum / recombinant protein / geratine particle / an easy and quick serodiagnostic method / monoclonal antibody

Research Abstract

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.