| Avian influenza (AI) is a severe disease devastating the stock-breeding and human health which is characterized by the blood poisoning of the respiratory system and the whole body of the bird. Since long ago it has outbreak in many countries and areas and has resulted in large economic loss. Therefore, people around the world attach much importance to it and do a lot of work on it. AI is readily regarded as the source of genetic variation of human influenza. The infection of H5N1 strain in Hong Kong in 1997, and H9N2 strain in both the mainland in Hong Kong gives AI more urgent public hearth concern..The diagnosis of AI is dependent on the isolation and characterization of the AIV, serology analysis, molecular diagnosis and electronic microscope technique. Serology tests include HA, HI, NIT, AGP, NT, ELISA and IFT. In 2004 a new method fluorescent RT-PCR is approved by State Plant Inspection and Quarantine Bureau as the new national standard to diagnose avian influenza. However, these methods have the disadvantages of time consuming and expensive lab equipment cost which make them not suitable for clinical instant diagnosis. Therefore, it is essential to establish a new fast diagnosis method. In this paper an instant method is successfully established to detect AI using Gold Immunochromatography Assay and will be very important.In this paper BALB/c mice are immunized with H5 strain derived AIV oil emulsion antigen vaccine from Haerbin Vet Institute for a long time. Then spleen cells from mouse wich have high high titer antibody are hybridized with SP2/0 bone marrow tumor cells with PEG-1500. The hybridization rate is 65.8%. An ELISA method is established to detect the McAb in the supernatant of the hybrid cell culture. Preliminary test shows a positive rate of 14.2%. After several sub-cloning and repetitive selection two hybrid cell lines are acquired namely 5A6 and 5C7 stably expressing anti-AIV McAb. Preliminary characterization of the two strains shows that they are both specifically against the common epitope of AIV. And they can both secrete high titer antibody above 1:1000 after repetitive frozen storage, thawed revival and generation, . The hybridoma cells are injected into the mice abdomen and seven days later ascites water is collected to purify the McAb by caprylic-sulfate ammonium method. The titers are above 1:32, 000. Preliminary biologic characterization of the McAbs shows that the McAbs are highly affinitive and specific. There are further proved to be IgGl k subtype by latex agglutination test. It is estimated that they will play important roles in the diagnosis of AI.After long period immunization of goats with H5 subtype oil emulsion antigen AI vaccine, high titer goat-anti-AIV polyclonal antibody is obtained which is proved by indirect ELISA and AGP. For the ELISA method the titer is above 1:100,000 and for the AGP method it is above 1:8 . With these... |
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