旋毛虫病人血清Western blot对成虫可溶性蛋白中早期诊断抗原的鉴定

Identification of soluble proteins from adult Trichinella spiralis worms in Western blots of sera from patients with trichinellosis and use of those antigens to diagnose trichinellosis in its early stages

目的鉴定旋毛虫成虫早期特异性诊断抗原。方法应用感染19d和35d的旋毛虫病患者血清,通过Western blot对旋毛虫成虫可溶性蛋白的抗原组分进行筛选和鉴定。结果 SDS-PAGE显示,旋毛虫成虫可溶性蛋白有27条蛋白带,分子质量单位分别为95、81.3、73.4、63.6、61.4、55、50.6、47、45.1、43.7、41.9、40.6、39、37.4、35.1、33.3、31、30.4、27.8、26.6、25.4、24、21、19.2、17.9、15.4和14.4ku。Western blot显示,成虫可溶性蛋白中的41.9ku和50.6ku蛋白与感染19d和35d的旋毛虫病患者血清反应率均为100%(15/15),与囊尾蚴病、裂头蚴病、并殖吸虫病、华支睾吸虫病患者血清及健康人血清均无交叉反应,与棘球蚴病和日本血吸虫病患者血清的交叉反应率分别为23.8%和44.44%。结论旋毛虫成虫可溶性蛋白中的41.9ku和50.6ku蛋白用于人体旋毛虫病早期诊断具有高度的敏感性和良好的特异性。

Objective To identify antigens from adult Trichinella spiralis worms in order to promptly and specifically diagnose trichinellosis. Methods Soluble proteins from adult T. spiralis worms were analyzed using SDS-PAGE and Western blots of sera from patients with trichinellosis. Results SDS-PAGE indicated that the soluble proteins had 27 protein bands with a molecular weight （MW） of 95, 81.3, 73.4, 63.6, 61.4, 55, 50.6, 47, 45.1, 43.7, 41.9, 40.6, 39, 37.4, 35. 1, 33.3, 31, 30.4, 27.8, 26.6, 25.4, 24, 21, 19.2, 17.9, 15.4, and 14.4 ku. Western blotting indicated that 2 soluble proteins with an MW of 41.9 and 50.6 ku from adult T. spiralis worms were recognized by serum samples from patients with trichinellosis 19 and 35 days post-infection. The 2 soluble proteins reacted with serum samples at a rate of 100 %（15/15 samples）. The 2 proteins were not recognized by serum samples from patients with cysticercosis, sparganosis, paragonimiasis, or clonorchiasis or by serum samples from healthy persons. However, the two proteins cross-reacted with serum samples from patients with echinococcosis at a rate of 23.8% （3/13） and with serum samples from patients with schistosomiasis at a rate of 44.44% （4/9）. Conclusion Soluble proteins with an MW of 41.9 ku and 50.6 ku from adult T. spiralis worms had high sensitivity and good specificity for early diagnosis of human trichinellosis.