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目的寻找曼氏裂头蚴排泄分泌(excretory-secretory,ES)物中的特异性诊断抗原。方法应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)与免疫印迹(Western blot)对裂头蚴ES抗原中的蛋白组分进行分析及鉴定,并与可溶性抗原进行比较。结果 SDS-PAGE显示,裂头蚴可溶性抗原有19条蛋白带,分子质量单位分别为134、129、124、108、100、80、70、56、47、45、43、40、38、36、35、32、27、241、6 ku;ES抗原有7条蛋白带,分子质量单位分别为66、53、42、36、29、24、20 ku。Western blot表明,可溶性抗原中的362、4 ku蛋白组分可被裂头蚴感染小鼠及裂头蚴病患者血清识别,其他蛋白组分则无抗原活性,或与其他寄生虫感染小鼠或寄生虫病患者血清有交叉反应;ES抗原中的36 ku蛋白组分只可被裂头蚴感染小鼠及裂头蚴病患者血清识别,而不与旋毛虫、血吸虫、弓形虫感染小鼠及正常小鼠血清产生交叉反应,与囊尾蚴病、棘球蚴病、并殖吸虫病、华支睾吸虫病、日本血吸虫病患者血清及健康人血清亦无交叉反应。结论裂头蚴ES抗原中的36 ku蛋白组分为裂头蚴的特异性抗原,可用于裂头蚴病的血清学诊断和流行病学调查。
Objective To search for specific diagnostic antigen in excretory-secretory (ES) Methods The protein components of ES antigen of the sporozoite were analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, and compared with the soluble antigen. Results SDS-PAGE showed that there were 19 protein bands in the soluble antigen of the spider larvae, with molecular mass units of 134, 129, 124, 108, 100, 80, 70, 56, 47, 45, 43, 40, 38, 36, 35, 32, 27, 241, 6 ku; ES antigen has seven protein bands, molecular mass units were 66,53,42,36,29,24,20 ku. Western blot showed that 362 and 4 ku protein components in the soluble antigen could be recognized by the sera of mice infected with sporozoite and in patients with sparganosis, while the other protein components had no antigen activity or were not associated with other parasitic mice or parasitic diseases Serum cross-reaction; ES antigen 36 ku protein components can only be infected with sporozoite mice and patients with sparganosis serum identification, but not with Trichinella spiralis, Schistosoma infection Toxoplasma infection and normal mouse serum cross-reaction, Cysticercosis and cysticercosis, echinococcosis, paragonimiasis, Clonorchis sinensis, schistosomiasis japonica serum and healthy serum no cross-reaction. Conclusion The 36 ku protein of the ES antigen of the splenocyte is a specific antigen of the sporozoite and can be used for serological diagnosis and epidemiological investigation of the sporozoite.