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首页> 外文期刊>Journal of Clinical Microbiology >Serodiagnosis of toxoplasmosis by using a recombinant form of the 54-kilodalton rhoptry antigen expressed in Escherichia coli.
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Serodiagnosis of toxoplasmosis by using a recombinant form of the 54-kilodalton rhoptry antigen expressed in Escherichia coli.

机译：弓形虫病的血清学诊断是通过使用重组形式的大肠杆菌中表达的54千达尔顿混种抗原来进行的。

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摘要

A 330-residue carboxy-terminal antigenic fragment of the Toxoplasma gondii 54-kDa rhoptry protein (ROP2) was expressed in Escherichia coli as a fusion polypeptide containing a 48-amino-acid sequence derived from phage lambda protein Cro and E. coli protein LacI followed by six consecutive histidyl residues. Metal chelate affinity chromatography provided an easy way to isolate the recombinant product in a highly purified form (> 95%). When this material was used to develop an immunoglobulin G (IgG) enzyme-linked immunosorbent assay for diagnosis of toxoplasmosis, the test reached a sensitivity of 89%. The sensitivity of the assay was similar whether the sera contained T. gondii-specific IgM or were devoid of such IgM. It was also found that ROP2 is a conserved antigen since antibodies against the recombinant antigen could be detected in mice experimentally infected with 11 independent T. gondii isolates originating from infected human tissues tested. Thus, the 54-kDa rhoptry antigen could advantageously complement other previously described T. gondii antigens for the development of more-sensitive and more-informative recombinant antigen-based tests for toxoplasmosis diagnosis.
机译：弓形虫54-kDa rhoptry蛋白（ROP2）的330个残基羧基末端抗原片段在大肠杆菌中表达为融合多肽，含有48个氨基酸的噬菌体λ蛋白Cro和大肠杆菌LacI蛋白。随后是六个连续的组氨酸残基。金属螯合亲和色谱法提供了一种简便的方法来分离高纯度形式（> 95％）的重组产物。当使用这种材料开发用于诊断弓形虫病的免疫球蛋白G（IgG）酶联免疫吸附测定时，该测试的灵敏度达到89％。无论血清中含有弓形虫特异性IgM还是不含此类IgM，测定的灵敏度均相似。还发现ROP2是一种保守的抗原，因为可以在用11种独立的刚地弓形虫分离株进行实验性感染的小鼠中检测到针对重组抗原的抗体，所述分离株来自受试人体组织。因此，54 kDa rhoptry抗原可以有利地补充其他先前描述的弓形虫T抗原，以开发用于弓形虫病诊断的更敏感，信息量更大的基于重组抗原的测试。

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