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目的:鉴别一起不明原因导致的突发性发热事件病原体。方法:提取样本中的RNA后,使用针对于甲型H1N1流感病毒相关的不同浓度引物、探针和DNA聚合酶缓冲液等作为反应成份,按照不同的扩增程序,利用逆转录PCR(RT-PCR)和实时荧光定量PCR(real-time PCR)方法分别进行特异性DNA片段的扩增,对扩增产物的电泳图片和扩增图谱进行分析。结果:经RT-PCR方法检测后,分别扩增出235、527、153和80 bp的特异性片段,实时荧光PCR扩增曲线中,出现了与阳性对照一致的扩增图谱。结论:此次事件的病原体为甲型H1N1流感病毒,定性本次事件为甲型H1N1流感暴发事件。
Purpose: To identify unexpected fever-causing pathogens caused by unknown causes. Methods: After extracting the RNA from the samples, different concentrations of primers, probes and DNA polymerase buffer for the Influenza A (H1N1) virus were used as reaction components. According to different amplification programs, reverse transcription PCR (RT- PCR and real-time PCR were used to amplify the specific DNA fragments respectively. The electrophoretic picture and amplification map of the amplified products were analyzed. Results: The specific fragments of 235, 527, 153 and 80 bp were amplified by RT-PCR respectively. The real-time fluorescent PCR amplification curve showed a consistent amplification pattern with the positive control. Conclusion: The pathogen of this incident is Influenza A (H1N1) virus. Qualitatively, this incident is an outbreak of Influenza A (H1N1).