| H5N1 subtype avian influenza began to attack the southeastern Asia since late 2003,and recently spread to Europe and Africa by the migratory birds.It caused fatal disease on avian and human.Long inspect results indicate that H5N1 subtype avian influenza virus may become the candidate for the next pandemic influenza because of its quick variation,causing fatal disease,and infecting human.Presently,the big problem is the lack of rapid diagnostic tests for influenza.The existing diagnostic methods can not meet the needs of monitoring the influenza epidemic situation.The purpose of this research is to develop a new rapid antigen detection test for HA of H5N1 avian influenza virus.Also,comprehensive evaluation will be done.Our lab has a great deal of experience in H5 McAbs preparation.With the help of phylogenetic analysis of H5N1 virus isolated from southeastern Asia recently,we know the evolution genesis of the HA1 gene in H5N1,then choose the high pathogenic strain "A/Ck/HK/Yu22/02" as the main immunogen for H5 McAbs preparation.At last,we get 476 H5 McAbs.and characterization of all the H5 McAbs was done.All of these work lay the solid groundwork for diagnostics development.Four McAbs(1A6,13H8,2F10,6B7) were choosed from the panel of H5 McAbs to establish a new rapid antigen detection test for HA of H5N1 avian influenza virus(H5-Dot) and its manufacturing technique.To evaluate the detection capability of H5-Dot,much work had been done.Results showed that both the sensitivity(85/85) and specificity(61/61) for virus strains in the allantoic fluid were 100%;the sensitivity and specificity for field samples was 77.3%(92/119) and 99.2% (20/2527).The detect limit of H5N1 virus strains in the allantoic fluid was 4x10~7copies(4x10~6-5xl0~8 ),analytical sensitivity was 25 times higher than QuickVue A+B,and 8 times higher than Directigen.These results suggest that H5-Dot is an acceptable detection test,and can play an important role on monitoring the influenza epidemic situation.
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