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目的研制一种可用于疫区现场快速诊断家畜日本血吸虫感染的胶体金免疫层析试纸条。方法设计重组蛋白G并在大肠杆菌中表达,得到重组蛋白。对重组蛋白G进行胶体金标记并制备金标垫,分别用可溶性虫卵抗原(SEA)和重组蛋白G划线,作为检测线和质控线。采用组装的试纸条检测标准阴、阳性BALB/c鼠与新西兰大白兔血清,以及粪检阴、阳性的绵羊与水牛血清,评估其灵敏度和特异度;利用组装的试纸条检测前后盘吸虫病病牛血清,评价其交叉反应性。结果成功构建了重组蛋白G的原核表达质粒并在大肠杆菌中表达。以重组蛋白G制备的试纸条可用于检测日本血吸虫感染BALB/c鼠、新西兰大白兔、水牛、绵羊血清抗体。该试纸条检测日本血吸虫感染小鼠与兔血清的灵敏度、特异度均为100%;检测绵羊血清的灵敏度为100%,特异度为88.46%;检测水牛血清的灵敏度为94.44%,特异度为100%;其与前后盘吸虫交叉反应率为5.88%。结论成功研制能检测多种家畜血吸虫病的胶体金免疫层析试纸条,其检测家畜血吸虫感染的灵敏度和特异度均较高。
OBJECTIVE: To develop a colloidal gold immunochromatographic strip for the rapid diagnosis of Schistosoma japonicum infection in livestock in the endemic area. Methods Recombinant protein G was designed and expressed in E. coli to obtain recombinant protein. The recombinant protein G was labeled with colloidal gold and the gold standard pad was prepared. The soluble egg antigen (SEA) and recombinant protein G were respectively scored as detection line and control line. Assess the sensitivity and specificity of standard negative and positive BALB / c mice and New Zealand white rabbits’ sera as well as feces and positive females and buffalo sera using the assembled test strips; Disease bovine serum, evaluate its cross-reactivity. Results The prokaryotic expression plasmid of recombinant protein G was successfully constructed and expressed in E. coli. Test strips prepared with recombinant protein G can be used to detect serum antibodies to BALB / c mice, New Zealand white rabbits, buffalo and sheep infected with Schistosoma japonicum. The sensitivity and specificity of the test strips for detection of Schistosoma japonicum-infected mice and rabbit serum were both 100%. The sensitivity and specificity of sheep serum were 100% and 88.46%, respectively. The sensitivity and specificity of detecting the serum of buffalo were 94.44% 100%; its cross-reaction rate with the former and the latter is 5.88%. Conclusion The colloidal gold immunochromatographic test strips capable of detecting various schistosomiasis strains were successfully developed. The sensitivity and specificity of detection of schistosoma infection in domestic animals were high.