肺炎衣原体43和53千道尔顿重组蛋白血清免疫印迹检测的评价

L. Campbell, S. Roberts, Shiuichiro Inoue, L. Kong, C. Kuo
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引用次数: 12

摘要

肺炎衣原体是呼吸道感染的常见病因。它也被证明与冠心病有关。据报道,在人类感染过程中经常被识别的两种蛋白质是分子质量为43和53 kDa的蛋白质。为了开发一种有用的替代微免疫荧光(micro-IF)测定的血清学试验,重组43-kDa和53-kDa衣原体特异性蛋白用斑点印迹和/或与标准微免疫荧光试验进行比较。从编码53-kDa蛋白的2个肺炎原体基因(CPn0809和CPn0980)和编码43-kDa蛋白的4个功能未知的肺炎原体基因(CPn0562、CPn0927、CPn0928和Cpn0929)的基因组序列信息中获得扩增引物,这些基因是衣原体特异性的,在沙体衣原体中未发现。CPn0809的53-kDa蛋白产物或n端18-kDa部分的特异性比任何43-kDa重组体都好,但灵敏度远低于micro-IF。相比之下,CPn0980编码的53-kDa蛋白被12个急性期血清中的11个(92%),46个慢性血清中的35个(76%),12个微if阴性血清(肺炎c和c。沙眼阴性),12例中1例(8%)C。肺炎阴性,沙眼衣原体阳性。因此,CPn0980编码的53-kDa蛋白可能用于c的血清诊断。肺炎感染。
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Evaluation of Chlamydia pneumoniae 43- and 53-Kilodalton Recombinant Proteins for Serodiagnosis by Western Blot
ABSTRACT Chlamydia pneumoniae is a common cause of respiratory infection. It has also been shown to be associated with coronary heart disease. Two proteins that have been reported to be recognized frequently during human infection are proteins having molecular masses of 43 and 53 kDa. In order to develop a useful alternative serological test to the microimmunofluorescence (micro-IF) assay, recombinant 43-kDa and 53-kDa chlamydia-specific proteins were evaluated in dot blot and/or for comparison to the standard micro-IF test. Primers for amplification were derived from genome sequence information for two C. pneumoniae genes (CPn0809 and CPn0980) encoding 53-kDa proteins and four C. pneumoniae genes (CPn0562, CPn0927, CPn0928, and Cpn0929) encoding 43-kDa proteins of unknown function, which wereChlamydia specific and not found in Chlamydia trachomatis. The 53-kDa protein product of CPn0809 or the N-terminal 18-kDa portion had better specificity than any of the 43-kDa recombinants but was much less sensitive than micro-IF. In contrast, the 53-kDa protein encoded by CPn0980 was recognized by 11 of 12 (92%) acute-phase sera, 35 of 46 (76%) chronic sera, 0 of 12 micro-IF-negative sera (C. pneumoniae andC. trachomatis negative), and 1 of 12 (8%)C. pneumoniae negative, C. trachomatis positive sera. Thus, it appears that the 53-kDa protein encoded by CPn0980 has potential use for serodiagnosis ofC. pneumoniae infection.
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