[梅毒螺旋体抗原TpN17及TpN44.5一个表位的克隆表达与临床应用]
[Cloning and expression of Treponema pallidum antigen TpN17 and an epitope of TpN44.5 and clinical application].
作者信息
Song Linghao, Xin Xiaolan, Mao Xia
机构信息
Suzhou Exit-entry Inspection and Quarantine Bureau, Suzhou, Jiangsu 215000, China.
出版信息
Zhonghua Nan Ke Xue. 2004 Dec;10(12):922-4.
OBJECTIVE
To investigate the cloning and expression of Treponema pallidum (TP) specific antigen TpN17 and an epitope of TpN44.5 and the clinical application of this fusion antigen.
METHODS
TpN17 gene was amplified from the genome of TP and an epitope of TpN44.5 was spliced to the 5' end of TpN17 gene. This modified TpN17 gene was cloned into the expression vector pGEX4T-2. The recombinant fusion antigen was purified by affinity chromatography and then an enzyme-linked immunosorbent assay (ELISA) kit was prepared. With this kit, the sera of 10 normal persons and 10 TP patients were tested.
RESULTS
The molecular weight of the purified fusion antigen was 45,000. Tested with ELISA, 10 serum samples of the TP patients were positive and another 10 of the normal persons were negative. ELISA equipped with the GST-epi-TpN17 antigen showed higher sensitivity and specificity as compared with routine methods.
CONCLUSION
The recombinant fusion TP specific antigen GST-epi-TpN17 was suitable for the preparation of ELISA kit in clinical examinations.
目的
研究梅毒螺旋体（TP）特异性抗原TpN17及TpN44.5一个表位的克隆、表达及其融合抗原的临床应用。
方法
从TP基因组中扩增TpN17基因，并将TpN44.5的一个表位连接到TpN17基因的5′端。将修饰后的TpN17基因克隆到表达载体pGEX4T-2中。通过亲和层析纯化重组融合抗原，然后制备酶联免疫吸附测定（ELISA）试剂盒。用该试剂盒检测10名正常人和10名TP患者的血清。
结果
纯化的融合抗原分子量为45000。ELISA检测显示，10份TP患者血清样本呈阳性，另外10份正常人血清样本呈阴性。与常规方法相比，配备GST-epi-TpN17抗原的ELISA具有更高的敏感性和特异性。
结论
重组融合TP特异性抗原GST-epi-TpN17适用于制备临床检测用ELISA试剂盒。
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