华支睾吸虫未脱脂和脱脂成虫抗原的免疫印迹分析

Western blot analysis on the soluble greasy and degrease adult worm antigens of Clonorchis sinensis

目的寻找华支睾吸虫成虫抗原中具有特异性诊断价值的抗原组分。方法应用SDS-PAGE和Westernblot分析华支睾吸虫成虫未脱脂、脱脂的可溶性抗原蛋白组分的血清学反应特征。结果未脱脂成虫抗原有14条蛋白带可与华支睾吸虫病病人血清反应,其中分子质量单位为180、170、100、67、41、37、35、322、6 ku是主带,180、170、37、35 ku条带还可与日本血吸虫病、卫氏并殖吸虫病病人血清发生交叉反应。脱脂成虫抗原有10条蛋白带可与华支睾吸虫病病人血清反应,其中分子质量单位为180、100、67、37、35、242、0 ku是主带,此外,180 ku带可与日本血吸虫病和卫氏并殖吸虫病病人血清发生交叉反应,78 ku带可以与血吸虫病病人血清发生交叉反应。结论华支睾吸虫成虫抗原经脱脂处理后可减少与日本血吸虫病和卫氏并殖吸虫病病人血清交叉反应的蛋白组分数量。华支睾吸虫未脱脂和脱脂成虫抗原中主要的特异性抗原组分分别是100、67、41、32、26 ku和100、67、37、35、242、0 ku,这些组分抗原可用于华支睾吸虫病的免疫诊断。

Objective To find out the valuable specific antigens for immunodiagnosis of clonorchiasis. Methods The greasy, degrease adult worm antigens of Clonorchis sinensis were separated by SDS PAGE and transferred on the nitrocellulose membrane, then were recognized by the patient＇s sera of clonorchiasis, schistosomiasis and paragonimiasis respectively. Results 14 protein bands were recognized by the sera of clonorchiasis in the greasy antigens of adult worms, among which the protein bands with relative molecular weight （MW） of 180, 170, 100, 67, 41, 37, 35, 32, 26 ku were main bands, in addition, the bands of 180, 170, 37, 35 ku could also cross react with sera of schistosomiasis and paragonimiasis, There were 10 proteins bands recognized by sera of clonorchiasis in degrease adult worm antigens of C. sinensis, among which the protein bands with MW of 180, 100, 67, 37, 35, 24, 20 ku were main bands, in addition, the protein bands of 180 ku could be recognized by the sera of schistosomiasis and paragonimiasis, the band of 78 ku could al so be recognized by the sera of schisotosomiasis. Conclusion The degreasing for C. sinensis adult worm antigen could reduce the numbers of the fraction antigens which could cross-react with sera of schistosomiasis and paragonimiasis. The fraction of 100, 67, 41, 32, 26 ku in the greasy antigen and 100, 67, 37, 35, 24, 20 ku in the degreased antigens of C.sinensis adult worms are the main specific antigens. It could be applied to the specific immunodiagnosis of clonorchiasis.