日本血吸虫感染兔尿液中具有诊断价值的循环抗原组分分析  

Analysis of valuable diagnostic circulating antigens in urine of rabbits infected by Schistosoma japonicum

作　　者：孙成松[1] 汪奇志[1] 尹晓梅[1] 汪峰峰[1] 刘晓明[1] 张世清[1] 汪天平[1] 雷家慧[2] 李雍龙[2] 

机构地区：[1]安徽省血吸虫病防治研究所,合肥市230061 [2]华中科技大学同济医学院寄生虫学教研室

出　　处：《热带病与寄生虫学》2013年第1期11-14,10,共5页Journal of Tropical Diseases and Parasitology

基　　金：国家863计划项目(No2006AA02A251)

摘　　要：目的分析日本血吸虫感染兔尿液中具有诊断价值的循环抗原(CAg)组分,为建立尿液CAg免疫学诊断方法奠定基础。方法十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析尿液中蛋白抗原组分;甘氨酸盐酸缓冲液(简称甘盐液)解离尿液中循环免疫复合物(CIC),通过抗日本血吸虫可溶性虫卵抗原(Solube egg antigen,SEA)的鸡卵黄免疫球蛋白(IgY),采用免疫印渍技术(Western blot)对解离后的CAg组分进行分析。结果 SDS-PAGE结果显示,正常兔尿液中未见明显蛋白条带:感染兔尿液在未作CIC解离前,有2条主带和3条次带,主带分子量为146kDa和60kDa,次带分子量为108kDa、72 kDa和67 kDa;而利用甘盐液解离感染兔尿液中的CIC后,仅见1条主带和1条次带,分子量分别为68kDa和86kDa。对甘盐液处理后的感染兔尿液作Western blot分析,结果显示其与日本血吸虫感染兔血清和抗日本血吸虫SEA特异性IgY抗体反应的蛋白条带均为68kDa,而与正常兔血清和免疫前IgY反应不产生条带。结论日本血吸虫感染兔尿液中存在具有诊断价值的CAg组分,有望建立基于IgY的尿液CAg免疫学诊断方法。Objective To analyze the valuable diagnostic circulating antigens(CAg) in urine of rabbits infected by Schistosoma japonicum and provide evidence for constructing immunological diagnostic methods of urine CAg.Methods Protein antigens of urine were analyzed by SDS-PAGE.Circulating immune complexes(CIC) in urine were dissociated by glycine-hydrochloride buffer solution and then detected by Western blot based on chicken egg yolk immunoglobulin(IgY) of anti-soluble egg antigens(SEA).Results SDS-PAGE showed that there was no obvious protein band in healthy rabbit urine,but there were two main bands with the molecular weight of 146kDa and 60kDa and three secondary bands of 108kDa,72kDa and 67kDa before dissociating CIC in infected rabbit urine.There were only 1 main band of 68kDa and 1 secondary band of86kDa after dissociating CIC.Western blot showed the same protein band of 68kDa of infected rabbit urine after dissociating CIC had intensive reaction with the infected rabbit serum and anti-SEA IgY,but no protein band reacted with heathy rabbit serum and IgY from unimmunized egg yolk.Conclusion There were valuable diagnostic CAg in urine of rabbits infected by Schistosoma japonicum and immunological diagnostic methods of CAg could be constructed based on IgY.

关 键 词：日本血吸虫 尿液循环抗原 卵黄免疫球蛋白 免疫印渍技术 

分 类 号：R53[医药卫生—内科学] R44[医药卫生—临床医学]