旋毛虫肌幼虫排泄分泌物中特异性诊断抗原的研究

Studies on Specific Diagnostic Antigens in Excretory-secretory Products from Trichinella spiralis Muscle Larvae

目的 寻找旋毛虫肌幼虫排泄分泌 (ES)物中的特异性诊断抗原。 方法 应用SDS PAGE和Western印迹对旋毛虫肌幼虫体外培养 18、3 0h后的ES抗原中的蛋白组分进行研究。 结果 旋毛虫肌幼虫培养 18、3 0h后得到的ES抗原组分大致相同 ,两种ES抗原中主要蛋白带的分子量为 112、110、10 8、97、5 3、49、45、42、3 5、2 3和 16kDa。18hES抗原中的 10 2、97、95和 5 3kDa以及 3 0hES抗原中的 5 3、49、45和 43kDa均与并殖吸虫病、华支睾吸虫病、日本血吸虫病及囊尾蚴病患者血清发生明显的交叉反应。ES抗原中的 2 3kDa蛋白组分只与旋毛虫感染的大鼠、小鼠及患者血清反应 ,而不与上述其它寄生虫感染者、正常大鼠和小鼠及正常人血清发生交叉反应。 结论 旋毛虫肌幼虫ES抗原中的 2 3kDa蛋白组分为旋毛虫肌幼虫的特异性抗原 ,可用于旋毛虫病的血清学诊断及血清流行病学调查。

Objective To find out the specific diagnostic antigens in excretory-secretory (ES) products from muscle larvae of Trichinella spiralis. Methods The ES antigens (ESA) of Trichinella spiralis muscle larvae cultured in vitro at 18 h and 30 h were analyzed by SDS-PAGE and Western blotting. Results At different times after cultivation, the protein components of ESA of T. spiralis muscle larvae were similar. SDS-PAGE revealed that the molecular weight(MW) of the major bands of 2 ES antigens were 112, 110, 108, 97, 53, 49, 45, 42, 35, 23 and 16 kDa. Western blotting showed that the protein bands with 102, 97, 95 and 53 kDa in 18 h ESA and the protein bands with 53, 49, 45 and 43 kDa in 30 h ESA cross-reacted with sera from the patients with paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis, respectively. The protein component with 23 kDa in ESA only reacted with sera from the rats and mice infected with T. spiralis and the patients with trichinellosis, but not reacted with sera from animals and patients infected with other parasites, and sera from normal rats, mice and persons. Conclusion The protein component with 23 kDa in T. spiralis ESA is the specific antigen of T. spiralis muscle larvae and it could be applied to the serodiagnosis and seroepidemiological survey of trichinellosis.