棘球蚴特异性抗原的蛋白质印迹分析

目的　寻找细粒棘球蚴 (Eg)和多房棘球蚴 (Em)特异性抗原组分用于免疫诊断。　方法　对Eg、Em的囊液、原头节、囊壁角质层、囊壁生发层及犬泡状带绦虫、犬豆状带绦虫、羊细颈囊尾蚴、猪囊尾蚴等 4种异源性绦虫 ,共14种粗抗原进行免疫学分析。采用蛋白质印迹试验 (Western blotting)、Genesnap和Genetool分析软件比较分析不同抗原蛋白条带分别与囊型棘球蚴病 (CE)及泡型棘球蚴病 (AE)患者血清反应的差异。　结果　 14种粗抗原中与CE、AE患者血清发生交叉反应的 11条蛋白条带相对分子质量 (Mr)为 130000、100000、94000、80000、75000、66000、62000、52000、38000、32000及 24000 ;与CE患者血清有特异性反应的 5条蛋白条带Mr为 41000、40000、22000、160 00和12000 ;与AE患者血清具有特异性反应的 8条蛋白条带Mr为 120000、109000、86000、59000、43000、28000、20000及 18000。　结论　确定了Eg和Em共有的交叉反应性抗原和具有进一步研究价值的特异性抗原组分 ,为进一步分离和鉴定具有诊断价值的特异性抗原提供了基础资料

Western Blotting Analysis of Specific Antigens from Different Components of Echinococcus metacestodes

Objective To analyze antigens for searching specific antigenic components for immunodiagnosis of echinococcosis. MethodsFourteen crude antigens from different tissues (cyst fluid, protoscoleces, laminated layer and germinal layer) of Echinococcus granulosus and E.multilocularis metacestodes and other 4 species of cestodes were analyzed by Western blotting. The differences of protein bands were compared for the 14 crude antigens by reacting with pooled sera from cystic echinococcosis (CE) and alveolar echinococcosis (AE) patients. Results Eleven protein bands from the antigens reacted nonspecifically with sera from both CE and AE patients were {Mr 130 000}, {100 000}, {94 000}, {80 000}, {75 000}, {66 000}, {62 000}, {52 000}, {38 000}, {32 000}, {24 000}. The highly specific protein bands recognized by AE sera were {Mr 120 000}, {109 000}, {86 000}, {59 000}, {43 000}, {28 000}, {20 000}, {18 000}, and by CE sera were {Mr 41 000}, {40 000}, {22 000}, {16 000} and {12 000}. Conclusion Different antigens shared by the two species of Echinococcus were examined and potential antigenic proteins specific for AE or CE sera were found, providing useful information for further identifying specific antigens for immunodiagnosis.