摘要
目的应用旋毛虫Ts21基因重组蛋白建立诊断人体旋毛虫病及动物旋毛虫感染的快速血清学方法。方法以胶体金标SPA、旋毛虫Ts21重组蛋白与肌幼虫排泄-分泌(excretory-secretory,ES)抗原包被硝酸纤维素膜(NCM),分别建立Ts21重组蛋白-斑点免疫金渗滤法(Ts21-dot immunogold-filtration assay,Ts21-DIGFA)与ES-DIGFA,对旋毛虫病及其他寄生虫病患者血清和感染动物血清进行检测,并与ELISA检测结果进行比较。结果Ts21-DIGFA检测旋毛虫病患者血清抗体的敏感性和特异性分别为94.73%和86.96%,ES-DIGFA的敏感性和特异性分别为89.47%和86.96%,差异无统计学意义(χ2敏感=0.368,χ2特异=0,P>0.05);Ts21-ELISA的敏感性和特异性分别为94.73%和94.20%,与Ts21-DIGFA相比差异无统计学意义(χ2敏感=0,χ2特异=1.272,P>0.05)。Ts21-DIGFA检测旋毛虫感染猪血清的敏感性和特异性分别为88.89%和100%,ES-DIGFA的敏感性和特异性分别为94.44%和95.00%,差异均无统计学意义(χ2敏感=0,χ2特异=0,P>0.05)。小鼠感染300条旋毛虫后4周Ts21-DIGFA的血清抗体检出率为100%(10/10);感染10条及5条旋毛虫后6周血清抗体检出率均为100%(10/10)。DIGFA可在3 min内肉眼观察结果,抗原包被后的NCM和金标SPA在4℃至少保存6个月。结论Ts21-DIGFA检测旋毛虫抗体具有较高的敏感性和特异性及良好的稳定性和重复性,可用于旋毛虫病的血清学诊断及血清流行病学调查。
[Abstract] Objective To establish a rapid serological method for diagnosis of human tricinellosis and animal Trichinella infection using the recombinant protein of Trichinella spiralis gene Ts21. Methods Staphylococcal protein A （SPA） was labeled with colloidal gold, the nitrocellulose membrane （NCM） was coated by recombinant Ts21 protein and excretory-secretory （ES） antigens of T. spiralis muscle larvae to establish the Ts21-dot immunogold-filtration assay （Ts21- DIGFA） and ES-DIGFA. Anti-Trichinella antibodies in sera from patients with trichinellosis or other parasitosis, and from animals infected with Trichinella or other parasites were assayed by DIGFA. The results of DIGFA were compared with those of ELISA. Results The sensitivity and specificity of Ts21-DIGFA to detect anti-Trichinella antibodies in sera from patients with trichinellosis was 94. 73% and 86.96%, respectively; those of ES-DIGFA was 89. 47% and 86.96%, respectively. The difference of sensitivity and specificity between recombinant Ts21 protein and ES antigen have no statistical significance （ x^2sensetivity = 0. 368, X^2 specificity = 0, P 〉 0. 05）. The sensitivity and specificity of Ts21-ELISA was 94.73 % and 94.20%, respectively; Those between two methods （Ts21-DIGFA and Ts21-ELISA） have no statistical significance （x^2sensetivity = 0. 368, X^2 specificity = 1. 272,P〉0.05）; When the sera from pigs infected T. spiralis were assayed, the sensitivity and specificity of Ts21-DIGFA was 88.89% and 100%, respectively; that of ES-DIGFA was 94.44% and 95.00%, respectively; the difference of sensitivity and specificity between recombinant Ts21 protein and ES antigen have no statistical significance（x^2sensetivity = 0. 368, X^2 specificity = 0, P〉0.05）. In mice infected with 300 T. spiralis larvae, the sera antibody positive rate detected by Ts21-DIGFA was 100% （10/10） at four weeks after infection. In mice infected with 10 and 5 larvae, the antibody positive rate detected by Ts21 DIGFA was also 100% （10/10
出处
《中国病原生物学杂志》
CSCD
2008年第11期824-827,共4页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.30471450)
河南省教育厅科技攻关计划项目(No.2008A310015)
关键词
旋毛虫
斑点免疫金渗滤法
重组蛋白
排泄分泌(ES)抗原
ELISA
Trichinella spiralis
dot immunogold-filtration assay
recombinant protein
excretory-secretory（ES） anti-gens
ELISA
