肺炎嗜衣原体菌体抗原的制备及应用

Preparation and use of Chlamydophila pneumoniae somatic antigen ELISA

目的制备肺炎嗜衣原体(Chlamydophila pneumoniae,Cpn)菌体抗原,纯化后建立ELISA方法,用于血清Cpn抗体的检测。方法应用进口Cpn毒株感染Hep-2细胞,利用吖啶橙染色和直接免疫荧光染色判断Cpn感染细胞,制备Cpn抗原。用初步纯化的Cpn抗原建立ELISA方法检测人血清特异性抗体。结果吖啶橙染色和直接免疫荧光检测显示,Cpn成功感染Hep-2细胞。提取Cpn抗原,用ELISA检测84份病人血清特异抗体,阳性率为52.4%,ELISA试剂盒法的阳性率为51.2%,差异无统计学意义(P>0.05)。用Cpn菌体抗原ELISA检测209份病人血清,阳性率59.3%(其中心血管疾病患者血清138份,阳性率58.0%;呼吸系统疾病患者血清71份,阳性率62.0%);检测84份健康者血清,阳性率2.4%,差异有统计学意义(P<0.05)。结论Cpn菌体抗原ELISA检测血清抗Cpn抗体可用于判断Cpn感染;病人血清Cpn抗体阳性表明,某些心血管和呼吸系统疾病患者的致病原因可能与肺炎嗜衣原体感染有关。

Objective To detect serum antibodies in patients infected with Chlamydophila pneumoniae （Cpn）, an EI.ISA technique was established after the preparation and purification of Cpn somatic antigen. Methods An imported Cpn strain was used to infect Hep-2 cells. The infected cells were identified through acridine orange staining and direct immunofluorescenee staining. A technique of purified Cpn somatic antigen ELISA was established for the detection of human serum specific antibody. Results The results of acridine orange staining and direct immunofluorescence staining showed that Hep-2 cells were successfully infected with Cpn. In the detection of Cpn from sera of 84 patients, Cpn somatic antigen ELISA had a positive detection rate of 52.4% and an ELISA kit had a similar rate of 51.2%, though the difference was not statistically significant （P〉0. 05）. The positive rate of Cpn detection was 59.3% in sera from 209 patients tested with Cpn somatic antigen ELISA （positive rates of Cpn detected in 138 patients with cardiovascular diseases and 71 patients with diseases of the respiratory system were 58.0% and 62.0%, respectively）. There was 2.4% positive rate of detection from sera of 84 healthy individuals, and the difference in the rate of detection for these individuals and patients was statistically significant （P（0. 05）. Conclusion Cpn somatic antigen ELISA to detect serum antibodies may prove useful in determining Cpn infection. Positivity for IgG antibody of patient sera indicated that some diseases of the cardiovascular and respiratory systems may be associated with Cpn infection.