曼氏裂头蚴排泄分泌物中特异性诊断抗原的研究

Specific diagnostic antigens in excretory-secretory products of sparganum mansoni

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 determine the specific diagnostic antigens in excretory-secretory（ES） products from plerocercoids（sparganum） of Spirometra mansoni.Methods The ES antigens of S.mansoni were analyzed by SDS-PAGE and Western blot,and the results were compared with those of soluble antigens.Results The results of SDS-PAGE showed that the soluble antigens had 19 protein bands with a molecular weight（MW） of 134,129,124,108,100,80,70,56,47,45,43,40,38,36,35,32,27,24,and 16 ku;ES antigens had 7 protein bands with a MW of 66,53,42,36,29,24,and 20 ku.The results of Western blot demonstrated that two protein bands with 36 ku and 24 ku in soluble antigens were only recognized by sera from sparganum-infected mice and patients with sparganosis,the other protein bands in soluble antigens had no antigenicity or cross-reacted with sera from mice infected with other parasites and patients with other parasitoses.The 36 ku protein component in ES antigens was recognized only by sera from sparganum-infected mice and patients with sparganosis and did not cross-react with sera from mice infected with T.spiralis,S.japonicum,or T.gondii or with sera from normal mice,from patients with cysticercosis,echinococcosis,paragonimiasis,clonorchiasis,or schistosomiasis,or with sera from healthy persons.Conclusion The 36 ku protein component in ES antigens of S.mansoni is a sparganum-specific antigen and can be used in the serodiagnosis and seroepidemiological study of sparganosis.