Rastawicki W, Jagielski M
Zakład Bakteriologii PZH w Warszawie.
Med Dosw Mikrobiol. 1998;50(3-4):259-67.
The Mycoplasma pneumoniae FH strain routinely used in our laboratory for over 25 years as antigen in serological tests, 2 reference M. pneumoniae strains from ATCC (29342 and M129) and 3 isolates of M. pneumoniae obtained in 1995 from pneumonia patients were compared by SDS-PAGE, complement fixation test (CFT) and by Western-immunoblotting against human and rabbit serum samples with high level of mycoplasmal antibodies. On SDS-PAGE all M. pneumoniae strains showed the same number of 23 polypeptides on the gel with identical molecular weights. The same strains on immunoblotting against human and rabbit serum samples showed six bands: 170, 89, 75, 55, 38 and 33 kDa with the strongest antibody staining in 170-(P1 protein) and 89-kDa bands. Because of its known antigenic relationships Mycoplasma genitalium was used for comparison. The pattern of M. genitalium proteins on SDS-PAGE was similar to pattern of M. pneumoniae but distinguishable. On immunoblotting six proteins of M. genitalium (135, 127, 110, 95, 75 and 45 kDa) reacted with human and rabbits immunoglobulins for M. pneumoniae antigens. Furthermore in complement fixation test both antigens, prepared from M. pneumoniae and M. genitalium, reacted as well with human and rabbit immunoglobulins for M. pneumoniae and with rabbit immunoglobulins for M. genitalium. These cross-reactions observed in serological techniques could give false positive results in routine diagnosis of M. pneumoniae infections. In such situations showing on immunoblott of presence in tested serum sample of antibodies to 170- and 89 kDa proteins could confirm M. pneumoniae infection.
在我们实验室中作为血清学检测抗原常规使用超过25年的肺炎支原体FH菌株、来自美国典型培养物保藏中心(ATCC)的2株肺炎支原体参考菌株(29342和M129)以及1995年从肺炎患者中分离得到的3株肺炎支原体,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、补体结合试验(CFT)以及针对含有高水平支原体抗体的人血清和兔血清样本进行的蛋白质免疫印迹法进行了比较。在SDS-PAGE上,所有肺炎支原体菌株在凝胶上均显示出相同数量的23种多肽,分子量相同。相同的菌株在针对人血清和兔血清样本的免疫印迹中显示出六条带:170、89、75、55、38和33 kDa,其中170 kDa(P1蛋白)和89 kDa带的抗体染色最强。由于已知生殖支原体的抗原关系,将其用于比较。生殖支原体蛋白质在SDS-PAGE上的图谱与肺炎支原体的图谱相似但可区分。在免疫印迹中,生殖支原体的六种蛋白质(135、127、110、95、75和45 kDa)与人及兔针对肺炎支原体抗原的免疫球蛋白发生反应。此外,在补体结合试验中,由肺炎支原体和生殖支原体制备的两种抗原,与人及兔针对肺炎支原体的免疫球蛋白以及兔针对生殖支原体的免疫球蛋白均发生反应。在血清学技术中观察到的这些交叉反应可能会在肺炎支原体感染的常规诊断中产生假阳性结果。在这种情况下,在免疫印迹中显示检测血清样本中存在针对170 kDa和89 kDa蛋白质的抗体可以确诊肺炎支原体感染。