| Avian influenza, caused by influenza A-virus, is a sort of infectious disease that is prevalentamong the wild birds and domestic fowls. The outburst of highly pathogenic avian influenzawill definitely result in great economic loss in the aviculture industry, and also it will endangerthe health of us human being. It is vital to guarantee the positive relevance ratio, control theavian influenza in time, and keep modifying the means of controlling the avian influenza for thedevelopment of the aviculture industry and preventing the prevalence of human influenza. Dueto the fact that the volumes of the samples that are tested are large, we need to enrich the viruswithin the samples if it is under the condition that the virus concentration is relatively low.Usually, the following two methods are adopted: the method of PEG precipitation and themethod of super centrifugation. The two methods mentioned above are easy to carry out.However, the method of PEG precipitation will hinder the reaction of the PCR and they will allweaken the toxicity of the flu virus, and meanwhile, the tedious operation will also weaken thetoxicity of the flu virus and cause the cross contamination among the samples. This researchestablishes a enrichment method named bio-magnetic bead which can enrich the avian flu viruswithin the liquid and easy to operate. By making use of this method, the time taken by testingwill be decreased, the testing efficiency be improved, the false negative rate be lowered, andwill make sure that the virus still has biological activity, which will promote the following workand further analysis.1.The establishment of Bio-magnetic Bead MethodFetuin is rich in α-2,3-and α-2,6-sialic acid that a the virus receptor of both human flu andavian flu. According to the theory of mutual reaction between virus and receptor, we couple thefetuin with the amination beads and carboxylation beads to make the bio-magnetic beads. Itshows that compared with the excitation of amido beads, which takes the5%glutaricdialdehyde as the activator for1hour, it will get more amount of fetuin combination by takingthe7.5%glutaric dialdehyde as the activator for each milligram, and the amount is8.12μg; eachmilligram will couple9.8μg fetuin by taking pH4.5MES（2-（N-morpholine）ethanesulfonicacid）as the excitation-buffer fluid to active the carboxyl beads, and will couple9.25μg fetuin iftaking pH4.5PBS（phosphate buffer）as the excitation-buffer fluid. After the reaction between the two bio-beads mentioned above and the allantoic fluid with H5N1subtype flu virus, weconclude that the two bio-beads can concentrate the H5N1subtype flu virus.2.The primary research on the property and application of the bio-beadsThe sensibility and specificity of the bio-beads’ concentration ability on H5N1subtype fluvirus, the bio-beads’ concentration on H1N1subtype flu virus, the virus’ quality of adsorptionand release, the biological activity of the enriched flu virus and the application of the bio-beadsto enrich the H5N1subtype flu virus with the sludge and the air are all studied in our research.The conclusion demonstrates that the amination bio-beads can enrich and detect108timesamount of virus diluents, and the carboxylation bio-beads1015times; the bio-beads cannotenrich the Newcastle disease I-series virus, instead, it can enrich the H1N1subtype flu virus; thebio-beads of the enrichment avian flu virus and release the enriched virus when the temperatureis37℃and the virus released virus is still with blood clotting activity and thus canmultiplication with the chick embryo; the bio-beads can also enrich the avian flu virus within thesludge and the air. |
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