重组泡球蚴主要表面抗原在棘球蚴病免疫诊断中的效果

目的 研究非融合表达重组抗原用于棘球蚴病免疫诊断的效果。方法 将泡球蚴主要表面抗原基因编码序列克隆于原核非融合表达载体pQE30（＋）并进行表达。以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS—PAGE）及免疫印迹（WB）试验对重组抗原鉴定。亲和层析纯化重组抗原，用于酶联免疫吸附法（ELISA）检测患者血清特异性抗体。结果 SDS—PAGE及WB分析显示泡球蚴主要表面抗原基因在大肠埃希菌内得到了高效非融合表达。以此重组抗原进行ELISA试验，检测棘球蚴病68例，阳性率达97．1％，检测囊虫等其他感染性疾病80例及健康人血清20份，阴性符合率达98％。结论 泡球蚴主要表面抗原基因在大肠埃希菌中获得了高效非融合表达，以该重组抗原建立的ELISA试验，对棘球蚴病诊断具有较高的敏感性和属特异性。

Effect of recombinant immunodominant major surface antigen of Echinococcus multilocularis in immunological assay for diagnosis of echinococcosis

Objective To evaluate effect of the non-fused recombinant antigen in immuno diagnosis of echinococcosis. Methods The coding sequence of the target gene was cloned into prokaryotic non-fusion expression vector pQE30 ( + ) and expressed in E. coli. The recombinant protein was identified by SDS-PAGE and Western Blot, purified with affinity chromatography and used in ELISA to detect specific antibody in patients' sera. Results SDS-PAGE and Western blot demonstrated that Echinococcus multilocularis immunodominant major surface antigen gene was highly expressed in E. coli. The result of ELISA showed that out of 68 echinococcosis sera, 66 samples (97.1% ) were detected positively by the assay. Only 2 samples (2% ) were positive in 100 sera from cases with other infective diseases including cysticercosis or healthy persons. The sensitivity and specificity of the assay were 97.1% and 98% respectively. Conclusion Echinococcus multilocularis immunodominant major surface antigen gene has been highly expressed in E. coli in non-fusion form. ELISA with the recombinant protein is highly sensitive and genus-specific for echinococcosis diagnosis.