Zheng J, Ding T, Chen Z, Fang H, Li H, Lu H, Wu Y
Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
Eur J Clin Microbiol Infect Dis. 2015 Jul;34(7):1319-26. doi: 10.1007/s10096-015-2343-8. Epub 2015 Mar 12.
Chlamydia pneumoniae causes diseases in humans, including community-acquired pneumonia, bronchitis, and sinusitis. It is also associated with atherosclerosis, coronary heart disease, and hyperlipidemia. In this study, we investigated novel materials with which to develop a sensitive and specific method to identify early C. pneumoniae infection, to allow more effective clinical treatment and prevention. We prepared novel monoclonal antibodies (mAbs) against a recombinant protein equivalent to the immunodominant region of chlamydial protease-like activity factor (CPAF) from C. pneumoniae. The mAbs specifically reacted with the endogenous CPAF antigen of the C. pneumoniae type strain in immunoblotting and indirect immunofluorescence (IIF) assays, but did not react with C. trachomatis type strains or genital secretions from patients with acute C. trachomatis infection. The mAb with the highest titer was used to develop a new IIF assay and enzyme-linked immunosorbent assay (ELISA) to detect the C. pneumoniae antigen in clinical specimens from child patients suspected of pneumonia. The sensitivity, specificity, and concordance rate of the mAb-based IIF and ELISA tests were compared with those of polymerase chain reaction (PCR). Our results show that these mAbs have excellent specificity and may be used to develop new screening tools for the diagnosis of early pediatric pneumonia.
肺炎衣原体可导致人类疾病，包括社区获得性肺炎、支气管炎和鼻窦炎。它还与动脉粥样硬化、冠心病和高脂血症有关。在本研究中，我们研究了新型材料，以开发一种灵敏且特异的方法来识别早期肺炎衣原体感染，从而实现更有效的临床治疗和预防。我们制备了针对与肺炎衣原体衣原体蛋白酶样活性因子（CPAF）免疫显性区域等效的重组蛋白的新型单克隆抗体（mAb）。在免疫印迹和间接免疫荧光（IIF）试验中，这些单克隆抗体与肺炎衣原体标准菌株的内源性CPAF抗原发生特异性反应，但不与沙眼衣原体标准菌株或急性沙眼衣原体感染患者的生殖道分泌物发生反应。使用效价最高的单克隆抗体开发了一种新的IIF试验和酶联免疫吸附试验（ELISA），以检测疑似肺炎儿童患者临床标本中的肺炎衣原体抗原。将基于单克隆抗体的IIF和ELISA试验的敏感性、特异性和符合率与聚合酶链反应（PCR）的进行了比较。我们的结果表明，这些单克隆抗体具有优异的特异性，可用于开发诊断早期儿童肺炎的新筛查工具。
