肺炎衣原体荧光PCR检测方法的建立和应用

目的采用Taqman探针技术,组建肺炎支原体荧光PCR基因检测方法。方法对GenBank登录的肺炎衣原体的基因序列进行生物信息学分析,针对保守区域设计引物和探针,组建实时荧光PCR检测方法。对来自本院的560份临床咽拭子样本和肺泡冲洗液样本进行检测。采用流感病毒、副流感病毒、呼吸道合胞病毒和腺病毒进行特异性评价。结果本实时荧光PCR方法对甲型流感病毒(甲型、乙型)、副流感病毒、呼吸道合胞病毒、呼吸道腺病毒无特异性反应。本院的临床样本检出26份,其中咽拭子9份,肺泡冲洗液17份。结论肺炎支原体荧光PCR检测方法可用于肺炎衣原体感染的辅助诊断。

Detection of Chlamydia pneumonia with Fluorescent Real-Time PCR

Objective To develop Taqman technology based PCR for the detection of Chlamydia pneumoniae. Method Conserved DNA region of Chlamydia pneumoniae from GenBank was selected for the designing of the primers and probe. 560 clinical samples from Guangzhou Nanfang Hospital in 2007 were then analyzed. The specificity of the designed primers were evaluated using influenza A virus, influenza B virus, para-influenza virus, respiratory syncytial virus and adenovirus. Result Non-specific amplification was not observed. Chlamydia pneumoniae was detected in 26 samples, in which 9 samples were obtained from throat swabs and 17 samples were from alveolus fluid. Conclusion The developed RT-PCR detection method may be used in the clinic diagnose of Chlamydia pneumoniae infection.