摘要
目的用基因工程方法表达嵌合的梅毒螺旋体优势表位抗原,建立检测血清梅毒抗体的双抗原夹心酶联免疫方法。方法通过计算机软件分析选择梅毒螺旋体优势抗原表位,用聚合酶链反应(PCR)扩增优势表位基因,构建了梅毒螺旋体多优势表位嵌合抗原(rTpN15-TpN17-TpN47)表达载体,转化宿主菌BL21(DE3)进行表达,亲和层析柱法纯化获得高纯度融合抗原,并用其建立检测梅毒抗体的DAS-EIA。结果表达的优势表位嵌合抗原具有很好的抗原性。用其建立的嵌合抗原DAS-EIA检测确诊的50份阳性和30份阴性血清,阳性检出率和阴性检出率都是100%。结论嵌合抗原DAS-EIA法具有比间接EIA和重组单抗原DAS-EIA更高的灵敏度和检出正确率,其检测水平已经达到国外TPHA的水平,该方法的建立为临床检测梅毒开辟了新的领域。
Objective Using the gene engineering to establish a new method which named double antigen sandwiched enzyme-linked immunosorbent assay（DAS-EIA） to detect the serum Syphilis antibody. Methods The dominant epitopes of Treponema pallidum were analyzed and picked out by using computer software. The recombinant multi-epitope chimeric antigen （ rTpN15- TpN17-TpN47 ） expression vector of Treponema Pallidum was constructed, and transformed into E. coli BL21 （ DE3 ）. Then, the an-tigen was highly expressed, purified and used for development of DAS-EIA consequently. Results 50 positive sera and 30 negative sera which had been diagnosed were detected. The sensitivity and specificity of EIA were 100 % （50/50） and 100 % （30/30） , respectively. Conclusion The result showed the DSA-ELISA is a method in the same level as TPHA and it will be a very potential method for detecting serum Syphilis antibody.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2006年第7期403-405,408,共4页
The Chinese Journal of Dermatovenereology
关键词
梅毒
嵌合优势表位抗原
双抗原夹心法
Syphilis
Recombinant Treponema pallidum Multi-epitope chimeric antigen
Double antigen sandwiched enzyme-linked immunosorbent assay
