鸡包涵体肝炎病毒DNA探针的制备及其斑点杂交方法的建立-预防兽医学专业论文.docxVIP

朱后顺鸡包涵体肝炎病毒探针的制备及其斑点杂交方法的建立Abstract 朱后顺鸡包涵体肝炎病毒探针的制备及其斑点杂交方法的建立 Abstract Chicken inclusion body hepatitis(IBH)is one kind of acute infectious diseases caused by the avian adenovirus．There are 1 2 serotypes related to the disease of avian adenovims．The infected avian always died suddenly,autopsy shows severe anemia,jaundice，hemorrhage， necrosis and enlargement of liver，it Can some times cause inclusion bodies within the liver cytoblast．The disease，also known as anemia syndrome．The target tissue of avian adenovirus is mainly liver and it could form inclusion bodies within the liver cytoblast．As it can lead to the decrease in production of poultry and Layer chicken，cause immunologic deficiency and thus contribute to the immune Failure as well as Secondary infection．As the damage caused by avian inclusion body hepatitis more and more serious day by day,the disease calls much attention from all aspect concerned．However,there is still not a way to detect the pathogen briefly，quickly and economically．Dot blot hybridization becomes one l(ind of ideal method because of its simple operation and cost contr01． Design a pair of primer according to the present reference and at the same time design another pair of primer to set as contrast．The second pair of primer is designed on the basis of the conserve sequence of Hexon gene from NCBI GenBank．Get the target production from the amplification of the liver tissue by PCR．Then，through the retrieve of the target production from the gelatin，purification of the nucleic acid，the PCR production Can be cloned into the T-easy vector and thus transfer into DH5 Q．From the operation above，we obtain the recombinant bacteria,select twelve clone for sequencing．The result of the sequence comparison shows that there are as many as ten strains of the clone are in concord winl the sequence of the serotype 8 of avian adenovirus．The homology is varying from 98％to 98．8％． Extract and restriction ellzyme of the recombinant plasmid to make sure the target