检测甲型H1N1流感病毒富集方法的初步探索

The Primary Exploration of Enrichment Methods for Influenza A (H1N1) Virus

目的探索甲型H1N1流感病毒的富集方法,提高流感病毒核酸检测的灵敏度。方法分别用ProteinA/G及H1N1特异性血清处理流感病毒样品,根据国家流感中心设计的H1N1亚型检测通用引物,采用反转录-聚合酶链反应(RT-PCR)方法,扩增产物为527bp,来评价病毒的富集效果。结果两种富集方法均能提高检测灵敏度,用ProteinA/G富集处理后检测灵敏度可达10-6,用H1N1特异性血清沉淀富集处理后灵敏度为10-5。结论初步建立的富集甲型H1N1流感病毒方法,有效提高了流感病毒RT-PCR检测的灵敏度,可用于甲型H1N1流感病毒的检测。

ObjectiveTo explore the enrichment methods for influenza A （H1N1） virus and improve the detection sensitivity of the virus nucleotide. MethodsThe influenza A （H1N1） virus samples were enriched by Protein A/G and the specific serum respectively.Universal primers of H1N1 subtype, which is designed by the National Influenza Center were applied in RT-PCR method to evaluate the effect of the virus enrichment. The RT-PCR product was 527bp. ResultsThese two enrichment methods both could improve the detection sensitivity. The detection sensitivity of Protein A/G and H1N1-specific serum enrichment methods could reach 10-6 and 10-5 respectively. ConclusionSuch established enrichment methods for influenza A （H1N1） virus could effectively improve the detection sensitivity of influenza viruses through RT-PCR and could be used to detect Influenza A （H1N1） virus.