肺炎衣原体及肺炎支原体临床血清学检测试剂的研发及应用

The Development and Application in Detection kit of Chlamydia and Mycoplasma Pneumonia in Serum of Clinical Children

目的通过实验获得工程重组肺炎支原体和肺炎衣原体的抗原,探讨其临床诊断价值;分析儿童肺炎支原体和肺炎衣原体的发病情况。方法采集医院收治的社区获得性肺炎患儿的血标本进行肺炎支原体和肺炎衣原体检测,并通过PCR方法得到肺炎衣原体的特异性抗原Cpn-MOMP和Cpn-CPAF和肺炎支原体P1-C和P1-B基因片段,应用工程重组抗原进行肺炎衣原体抗原特异性及灵敏度检测和肺炎支原体的灵敏度实验。结果肺炎支原体及肺炎衣原体感染的阳性率随着年龄的增长而增加,肺炎衣原体冬季发病率较高。PCR方法可以得到完整的Cpn-MOMP和Cpn-CPAF基因片段,且Cpn-MOMP重组抗原产量高特异性较好。重组肺炎支原体P1-C和P1-B抗原产量较满意。结论肺炎支原体及肺炎衣原体感染阳性率逐年升高,通过重组的Cpn-MOMP和Cpn-CPAF抗原可用于肺炎衣原体感染的血清学检测,具有临床应用前景。

Objective Study on the diagnosis value in recombinant antigen of chlamydia pneumonia and mycoplasma pneumonia. The relationship of morbidity of chlamydia pneumonia, mycoplasma pneumonia was analyzed with ages, seasons etc. Methods Blood sample were collected from patients of Community acquired pneumonia in 2013. Chlamydia pneumonia and mycoplasma pneumonia were detected by testing of antibodies in serum of inpatients. Gene fragments of Cpn-MOMP and Cpn-CPAF were obtained by PCR and the sensitivity and specificity of which were analyzed by using recombinant antigen. Results The infection rates of chlamydia pneumonia and mycoplasma pneumonia increased, according with the age, and the morbidity of chlamydia was higher in winter. Complete gene fragments of Cpn-MOMP and Cpn-CPAF were obtained by PCR, the output and specificity of recombinant antigen of Cpn- MOMP were better. The output of recombinant antigen of Cpn-CPAF was high. The outputs of recombinant antigens of gene fragments P1-C and P1-B of mycoplasma pneumonia were high. Conclusion The infection rates of chlamydia pneumonia and mycoplasma pneumonia increased in recent years. The recombinant antigen of Cpn-MOMP and Cpn-CPAF can be applied in serum detection of chlamydia pneumonia infection while the detection method of mycoplasma pneumonia needs further investigation.