中国寄生虫学与寄生虫病杂志 ›› 1998, Vol. 16 ›› Issue (3): 201-203.

• 论著 • 上一篇    下一篇

western blot 法诊断囊虫病的应用研究

王敏; 郭瀛军; 陈蕊雯; 徐之杰; 孙树汉   

  1. 哈尔滨医科大学寄生虫学教研室; 第二军医大学分子遗传学教研室
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-06-30 发布日期:1998-06-30

APPLICATION OF IMPROVED WESTERN BLOT METHOD IN DIAGNOSIS OF CYSTICERCOSIS

Wang Min 1; Guo Yingjun 2; Chen Ruiwen 2; Xu Zhijie 1; Sun Shuhan 2   

  1. 1 Department of Parasitology, Harbin University of Medical Sciences, Harbin 150086 2 Department of Molecular Genetics, Second Military MedicaI University
  • Received:1900-01-01 Revised:1900-01-01 Online:1998-06-30 Published:1998-06-30

摘要: 目的:探讨四种猪囊尾蚴(Cysticercuscelulosae)特异性抗原cC1、cC2、cP1、cH1(分子量分别为28kDa、18kDa、14kDa、34kDa)混合应用诊断人囊虫病的价值。方法:以猪囊尾蚴cDNA表达文库中筛选出的β-半乳糖苷酶-猪囊虫特异性抗原cDNA编码的融合蛋白(fusionproteins,FP)为抗原,作简化的Westernblot(improvedWesternblot,IWB)分析,检测107例囊虫病、40例华支睾吸虫病、24例包虫病和34例健康人血清对融合蛋白的反应。同时应用粗制抗原(crudeanti-gen,CA)进行ELISA、IHA对比检测。结果:在简化Westernblot检测中,FP被107例囊虫病患者血清中94例(87.9%)血清所识别,与华支睾吸虫病、包虫病患者及健康人血清无交叉反应,特异性为100%。以粗制抗原作ELISA、IHA检测囊虫病人血清中的IgG抗体阳性率分别为84.1%和74.8%,与华支睾吸虫病患者血清存在交叉反应,假阳性率分别为2.5%、12.5%;与包虫病患者血清存在交叉反应,假阳性率分别为8.3%、16.7%;与健康人血清也存在交叉反应, 假阳性率分别为8. 8%、11. 8% 。结论: β-半乳糖苷酶2猪囊尾蚴FP 诊断囊虫病具有较高敏感性和高度的特异性。

关键词: 囊尾蚴病, 免疫诊断, 融合蛋白, 简化Western blot

Abstract: AIM:To evaluate the diagnostic value of four specific antigens from cDNA of
Cysticercus cellulosae . cC1,cC2,and cP1 and cH1 ( 28 kDa, 18 kDa, 14 kDa and 34 kDa ), mixing in equal proportions for the diagnosis of cysticercosis. METHODS:Taking the FP (fusion proteins) as antigen to make IWB (improved Western blot ) analysis basing on the detection of antibody
responses against FP,and making ELISA / IHA crude antigen(CA) analysis.They were evaluated
comparatively while using l07 infected sera of cysticerco sis cases, 40 infected sera of clonorchiasis cases, 24 infected sera of ech inococcosis cases and 34 sera of healthy persons. The FP are encoded by cDNAs of β-galacto sidase-specific antigens of Cysticercus cellulosae isolated from the cDNA library. RESULTS: 94 (87. 9% ) sera from l07 cysticercosis cases recognized FP in IWB and could not cross-react with the sera of echinococcosis cases, clonorchiasis cases and healthy persons, the specific rates were 100%, whereas ELISA, IHA using CA were 84. 1%and 74. 8%, respectively and could cross-react with the sera of echinococcosis cases, the false positive rates were 2. 5% and 12. 5% respectively; CA 2EL ISA/IHA could cross-react with the sera of clonorchiasis patients, the false positive rates were 8.3% and 16. 7%, respectively; and they could also cross-react with the sera of healthy persons, the false positive rates were 8. 8% and 11. 8% , respectively. CONCLUSION: The recombinant FP used in the immunodiagnosis of cysticercosis is specific and sensitive.

Key words: Cysticercosis, immunodiagnosis, fusion protein, improved Western blot