【摘要】 目的:构建人IA-2基因不同区段原核表达载体,诱导表达获得重组蛋白,并初步验证其在1型糖尿病蛋白酪氨酸磷酸酶自身抗体检测中的价值。方法:用RT-PCR方法调取目的基因,构建相应的原核表达质粒,转化大肠杆菌HB101,诱导表达获得纯化重组蛋白;以重组蛋白为包被抗原,初步建立检测蛋白酪氨酸磷酸酶自身抗体的ELISA方法,评价各片段在1型糖尿病诊断中的价值。结果:获得了2种可被1型糖尿病患者血清识别的重组人蛋白酪氨酸磷酸酶抗原区段IA-2(601～979)和IA-2(683～979),检测敏感性和特异性相当,但IA-2(683～979)检测的阳性D450nm值明显高于IA-2(601～979),成为首选的抗原区段。结论:所选重组人IA-2(683～979)抗原区段具有良好的抗原性,可作为1型糖尿病患者辅助诊断试剂的候选抗原。更多还原

【Abstract】 Objective:To obtain two different recombinant fragments of human protein tyrosine phosphatase-like protein IA-2,and to evaluate their diagnostic application for type 1 diabetes mellitus.Methods:The two DNA fragments of the IA-2 were obtained by RT-PCR and nested-PCR.The corresponding prokaryotic expression vector pBVIL1/IA-2 was constructed and transformed into E.coli HB101 to induce the expression.The two antigen fragments were used as the coating antigens,and the ELISA was established for the detection of the IA-2 autoantibody.Results:The two fragments of human IA-2 both reacted with the serum of type 1 diabetes mellitus patients. The sensitivity and the specificity of the IA-2(683～979)fragment were similar to the IA-2(601～979)fragment,but the D450nm value read from IA-2(683～979)fragment was higher than that from IA-2(601～979)fragment.Thus,the I-A-2(683～979)fragment became the preferred antigen.Conclusion:Because of the prior antigenicity of the recombinant human IA-2(683～979),it could be the candidate antigen for developing the diagnostic reagent for type 1 diabetes mellitus.更多还原