兰州市首起甲型H1N1流感暴发的核酸检测  

Nucleic acid assay techniques for pathogen of first outbreak H1N1 influenza in Lanzhou

作　　者：刘萍[1] 赵英杰[1] 马汉平[1] 罗军德[1] 罗晓梅[1] 施文静[1] 

机构地区：[1]兰州市疾病预防控制中心,兰州730030

出　　处：《中国卫生检验杂志》2010年第2期312-314,共3页Chinese Journal of Health Laboratory Technology

摘　　要：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。Objictive：To identify the pathogen that lead to an unexpected public fever accident.Methods：The reaction mixture is to utilize primers,probes,Taq polymerase and buffer solution with particular concentration designed for H1N1 virus.According to an amplification procedure after the total RNA was purified,two different methods were operated to detect the presence of target product by different size fragments and specific amplified curves respectively.Results：The target size fragments including 235,527,153,80 bp are obtained by RT-PCR.From the real-time PCR amplification plot,special amplified curves appeared complete similarity with a positive template control.Conclusion：The outbreak accident pathogen is revealed for the presence of H1N1 influenza virus.

关 键 词：甲型H1N1流感 暴发 PCR 

分 类 号：R511.7[医药卫生—内科学]