Development of the rapid diagnosis test for avian influenza viruses

由变异禽流感病毒衍生的大流行性流感的爆发一直是世界上令人担忧的问题。当流感大流行爆发时，免疫层析快速诊断试验是区分流感大流行与季节性流感和其他发热性疾病必不可少的。为了建立快速诊断试验，制备了与位于A/Turkey/安大略/7732/66（H5 N9）（免疫原）的NP的1/3 N-末端的表位反应的单克隆抗体4 E3，并将其用于免疫层析（夹心法）的固相和有色胶乳缀合抗体。该快速检测试剂盒在10分钟内至少检测到4 × 10^5个病灶形成单位（FFU）/ml的A/Crow/京都/04（H5 N1），并分别检测到3.4 × 10^5、5.3 × 10^5、1.1 × 10^5个FFU/ml的H5 N2、H5 N3、H5 N9病毒。除H5株病毒外，其余15株（H3 ~ H15）均与该病毒呈阳性反应。另一方面，它不与19种人流感病毒（A和B型）、22种人流感病毒（B型）、22种人流感病毒（C型）、22种人流感病毒（B型） 关于我们 其他呼吸道病毒或24种细菌。此外，快速检测法检测到所有10株AIV 2004年在泰国从10名患者中分离到H5 N1，Z型，进化枝1，并检测到1株AIV（H5 N1，可能是进化枝2-1），从2头病猪分离的2株H5 N1病毒中的2株，2006年从印尼7只死鸡中分离到7株H5 N1病毒。目前，AIV感染患者鼻咽部的H5 N1病毒载量似乎不像季节性流感那样高。然而，一旦大流行性流感病毒出现，这种病毒在人类中具有很强的传染性，病毒可以在鼻咽区域充分传播，达到季节性流感的水平。这些结果表明，该快速诊断试验具有足够的灵敏度、特异性、抗原识别的稳定性和快速性能。这种检测应显示出作为快速遏制和分诊的有力工具的巨大能力，特别是在大流行性流感的早期阶段。少

The outbreak of a pandemic influenza derived from a mutated avian influenza virus has been an apprehensive issue in the world. When the pandemic influenza broke out, a rapid diagnosis test by immunochromatography should be indispensable to distinguish the pandemic from the seasonal influenza and other febrile illness. To establish the rapid diagnosis test a monoclonal antibody, 4E3, which reacts with an epitope located in 1/3 of N-terminus of NP of A/Turkey/Ontario/7732/66(H5N9) (immunogen) was produced and used for both solid-phase and colored latex-conjugated antibody of an immunochromatography(sandwich method). This rapid test detected at least 4x10^5 focus forming unit(FFU)/ml of A/Crow/Kyoto/04(H5N1) in 10 min, and it detected 3.4x10^5, 5.3x10^5, 1.1x10^5FFU/ml of an H5N2, H5N3, H5N9 virus, respectively. It also reacted with 15 strains(H3〜H15) of avian influenza viruses(AIV) except H5 viruses. On the other hand it neither reacted with 19 human influenza viruses(A and B type), 22 o … More ther respiratory viruses or 24 bacteria. Furthermore, the rapid test detected all 10 strains of AIV(H5N1, Z type, clade 1) isolated from 10 patients in 2004 in Thailand and it also detected 1 of 1 strain of AIV(H5N1, probably clade 2-1) isolated from one patient, 2 of 2 strains of H5N1viruses isolated from 2 ill pigs, and 7 of 7 strains of H5N1 viruses isolated from 7 dead chickens in 2006 in Indonesia. At present H5N1 viral load at nasopharyngeal region in patients with AIV infections seems not high as in case of seasonal influenza. However, once the pandemic influenza viruses emerged which is very contagious among humans, the viruses could propagate sufficiently in the nasopharyngeal region to the level of seasonal influenza. These results reveal that this rapid diagnosis test has enough sensitivity, specificity, stability for antigen recognition and rapid performance. This test should display great ability as a powerful tool for the rapid containment and triage especially in the early-phase of the pandemic influenza. Less