日本血吸虫成虫及虫卵可溶性抗原的早期诊断分子筛选

为获取有效的日本血吸虫病早期诊断靶抗原分子。以感染前和感染后2周、4周、6周兔混合血清以及急性、慢性日本血吸虫病人和正常人血清,用Westernblot对日本血吸虫成虫可溶性抗原(AWA)和虫卵可溶性抗原(SEA)进行了全面的分析与筛选。SEA140kDa和AWA54kDa、21kDa、20kDa分子出现最早,能被感染后2周兔血清所识别;SEA69kDa、50kDa、45kDa、38kDa和AWA72kDa、45kDa、34kDa、15kDa相继能被感染后4周兔血清所识别;其中SEA140kDa、50kDa、45kDa、38kDa和AWA34kDa、21kDa、54kDa、15kDa与病人血清反应同6周兔血清反应效果相仿,均为免疫反应主带,且在SDS-PAGE上有对应的蛋白主带,具有潜在的早期诊断价值。进一步对SEA进行抗体类型的分析,发现SEA能刺激机体产生IgG、IgM和IgA反应,其中SEA50kDa、45kDa、38kDa三个抗原分子均能被IgG、IgM、IgA三种抗体所识别,且均为反应主带;SEA140kDa以IgM反应带最深;SEA100kDa反应带仅出现于病人血清,以IgM反应最强,且其与IgM反应带在急性病人血清明显深于慢性病人血清,表明针对IgM的SEA100kDa分子也具有一定的早期诊断和区分病程的价值。SEA140kDa、50kDa、45kDa、38kDa和AWA34kDa、21kDa、54kDa、15kDa和针对IgM的SEA100kDa分子是具有潜在早期诊断价值的优势靶抗原分子。

SCREENING OF EARLY DIAGNOSTIC MOLECULES OF THE SOLUBLE ADULT WORM ANTIGEN (AWA) AND SOLUBLE EGG ANTIGEN (SEA) OF SCHISTOSOMA JAPONICUM

In order to obtain the effective early target antigen molecules of Schistosoma japonicum for immunodiagnosis. The soluble adult worm antigen (AWA) and soluble egg antigen (SEA) of Schistosoma japonicum have been screened and analyzed using Western blot with sera of rabbits before and 2W,4W,6W after infection and sera of acute,chronic Schistosomiasis patients and normal human beings. SEA 140kDa and AWA 54,21,20kDa molecules appeared earliest,and could be recognized by rabbits sera after 2W infection. Reactions to SEA 69,50,45,38kDa and AWA 72,45,34,15kDa antigens appeared 4W post infection. Among these early antigens,SEA 140,50,45,38kDa and AWA 34,21,54,15kDa molecules reacted with both Schistosomiasis patients sera and rabbits sera after 6W infection. These eight antigens caused strong immune response and showed themselves to be main protein bands by SDS PAGE. It suggested that these eight molecules would be potential early antigens for immunodiagnosis. Further experiments to analysis immunoglobulin type response against SEA revealed that SEA could stimulate body to produce IgG and IgM and IgA antibodies. SEA 50,45,38kDa molecules induced strong IgG and IgM and IgA responses while SEA 140kDa predominantly resulted in IgM respones. Only reacted with patients sera and resulted in predominant IgM response,SEA 100kDa band against IgM of acute Schistosomiasis patients sera was obvious darker than that of chronic Schistosomiasis patients sera. This showed that the anti IgM SEA 100kDa antigen might have the potential early immunodiagnostic value. SEA 140,50,45,38kDa and AWA 34,21,54,15kDa and Anti IgM SEA 100kDa of Schistosoma japonicum were the potential early diagnostic molecules.