间接ELISA检测犊牛轮状病毒抗体方法的建立及其疫苗的初步研制-基础兽医专业论文.docxVIP

ABSTRACTBased ABSTRACT Based the basis of laboratory conditions，by ordinary high-speed centrifuges instead of low·temperature higll—speed centrifuges，Coke Dicthyl(DEPC)water treatment system for plastic， rubber-made and the general improvement measures，such electrophoresis tank，the extraction of higher rotavims RNA，the law also applies to other sexually transmitted diseases of the original RNA extraction． Although this law in small amount of RNA degradation，but does not affect the results，this relatively weak conditions for the grass-rooks epidemic prevention and supervision units，still has good feasibility and significance of the promotion． This study was collection from calf diarrhea outbreak in Shandong．RT-PCR detected the specific purpose ofthezone，SDSdetected rotavirusinll specificzones．Agroupconfirmedthatthe mtavirus． Cattle NCDV standard strain of rotavirus antigen，its high for the preparation of anti-serum horseradish peroxidase labeled goat anti-IgG the anti—cattle，the of orthogonal test with phalanx of detecting calf Bovine rotavirus antibodies indirect ELISA method．The experimental of orthogonal experiment with square indirect ELISA method to determine the best conditions：coating antigen 51．5 ng， hole，150 times the serum dilution，dilution ELISA antibody 2500，the detection of bovine rotavirus sensitivity of 36．78 ng． This experiment were aimed to develop the most—effective rotavirus vaccine for reduce economic losses of cattle．The first was approved virus TCID50，virulence of the virus in mice，specific tests， immunogenicity trial，and purity check．Preparation of the vaccine sterile test and safety test results were meet the standards，werg immune calves and pregnant cows，respectively．The trial of the virus to accord with the standard，bovine rotavirus vaccine were immune calves and pregnant cows，respectively．The control group and the immune group lgG antibody levels were significantly of calves；the control group and immune group of Pregnant cow’s milk-IgG anti