Indirect Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Reactive with a Recombinant Protein Expressed from the Gene Encoding the 116-Kilodalton Protein of Mycoplasma pneumoniae

Serology remains the method of choice for laboratory diagnosis of Mycoplasma pneumoniae infection. Currently available serological tests employ complex cellular fractions of M. pneumoniae as antigen. To improve the specificity of M. pneumoniae diagnosis, a recombinant protein was assessed as a serodiagnostic reagent. A panel of recombinant proteins were expressed from a cloned M. pneumoniae gene that encodes a 116-kDa surface protein antigen. The recombinant proteins were assessed for reactivity with patient sera and the most antigenic was further assessed for its serodiagnostic potential by indirect enzyme-linked immunosorbent assay (ELISA). The ELISA based on the recombinant protein was equivalent in sensitivity to the commercial test (Serodia Myco II; Fujirebio Inc.) to which it was compared. Southern and Western blotting data suggested that the recombinant protein derived from the 116-kDa protein of M. pneumoniae could provide a species-specific diagnostic tool, although further assessment is required.

血清学仍是肺炎支原体（Mycoplasma pneumoniae）感染实验室诊断的首选方法。当前可用的血清学检测以肺炎支原体的复合细胞组分作为抗原。为提升肺炎支原体诊断的特异性，本研究评估了一种重组蛋白作为血清诊断试剂。研究人员从编码116 kDa表面蛋白抗原的克隆化肺炎支原体基因中表达了一组重组蛋白，通过患者血清检测这些重组蛋白的反应性，并选取抗原性最强的重组蛋白，通过间接酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）进一步评估其血清诊断潜力。基于该重组蛋白的ELISA检测灵敏度与对照的商业化检测试剂盒（赛罗迪亚支原体II（Serodia Myco II）；富士瑞必欧株式会社）相当。Southern印迹法与Western印迹法结果显示，源自肺炎支原体116 kDa蛋白的重组蛋白可作为一种种特异性诊断工具，不过仍需开展进一步评估。