【摘要】 目的　用基因工程方法表达梅毒螺旋体的TmpA重组抗原 ,建立检测梅毒螺旋体抗体酶联免疫试验 (EIA)。方法　用聚合酶链反应 (PCR)扩增梅毒螺旋体TmpA基因 ,克隆到原核表达载体 pQE 30中表达 ,用亲和层析柱法纯化该重组抗原 ,并用其建立检测梅毒螺旋体抗体的EIA。 结果　表达的重组抗原相对分子质量约为 390 0 0 ,具有良好的抗原性。用其建立的EIA检测 10份阳性参比血清 ,均为阳性 ,灵敏度为 10 0 % ;检测 2 0份阴性参比血清 ,均为阴性 ,特异性为 10 0 %。检测12份Ⅰ期梅毒 (早期感染 )和 2 4份Ⅱ期和晚期梅毒患者血清 ,阳性率分别为 91.6 7% (11/ 12 )和 10 0 %(2 4/ 2 4)。结论　用该重组抗原建立的EIA可检出 97.2 % (35 / 36 )以上梅毒患者 ,仅 2 .8% (1/ 36 )漏检。为进一步提高该试剂的灵敏度 ,可能需增加梅毒螺旋体的其它重组抗原更多还原

【Abstract】 Objective To express TmpA recombinant antigen of Treponema pallidum in E.coli and to develop an Enzyme linked Immunoassay (EIA) based on the recombinant antigen for diagnosing syphilis. Methods The target TmpA gene was amplified by polymerase chain reaction (PCR). The PCR products of the gene were inserted into pBluescript T vector, and then expressed in E.coli, using pQE 30 system. Then the recombinant antigen was purified by an affinity chromatography and used for the development of EIA. Results The antigenicity of the recombinant antigen was identified by western blotting (WB) and EIA. The sensitivity and specificity of EIA were 100%(10/10) and 100% (20/20), respectively. The positive rates of anti TmpA antibodies were 91.67% (11/12) for the patients with Ⅰ phase of syphilis and 100% for the patients of Ⅱ and late stage of the disease. Conclusions The TmpA recombinant protein can be used to diagnose syphilis since 97.2% (35/36) of patients with syphilis were positive for the anti TmpA antibody by EIA.更多还原