western blot 法诊断囊虫病的应用研究

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。

APPLICATION OF IMPROVED WESTERN BLOT METHOD IN DIAGNOSIS OF CYSTICERCOSIS

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 107 infected sera of cysticercosis cases, 40 infected sera of clonorchiasis cases, 24 infected sera of echinococcosis cases and 34 sera of healthy persons. The FP are encoded by cDNAs of beta-galactosidase-specific antigens of Cysticercus cellulosae isolated from the cDNA library. RESULTS: 94(87.9%) sera from 107 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-ELISA/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.