Establishment Of The Indirect ELISA To Detection BRV Antibody Methods And The BRV Vaccine Development

Based on the basis of laboratory conditions, by an ordinary high-speed centrifuges instead of low-temperature high-speed centrifuges, Coke Diethyl (DEPC) water treatment system for plastic, rubber-made and the general improvement measures, such as electrophoresis tank, the extraction of higher rotavirus RNA, the law also applies to other sexually transmitted diseases of the original RNA extraction. Although this law in a small amount of RNA degradation, but does not affect the results, this relatively weak conditions for the grass-roots 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 of the zone, SDS-PAGE detected rotavirus in 11 specific zones. A group confirmed that the rotavirus.Cattle NCDV as a standard strain of rotavirus antigen, its high for the preparation of an anti-serum as, horseradish peroxidase labeled goat anti-IgG as the anti-cattle, the use of orthogonal test with a phalanx of detecting calf Bovine rotavirus antibodies indirect ELISA method. The experimental use 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, were 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 IgG antibody levels were significantly of calves; the control group and immune group of Pregnant cow's milk-IgG antibody titers were not significantly different. Analyze the failure of immune pregnant cow were vaccine may be less, the less time of immune, the feeding environmental impact and other reasons. The experimental has proved that feasibility of bovine rotavirus vaccine immune calves and lied the foundation of prevention of rotavirus diarrhea.vaccine immune calves and laid the foundation of prevention of rotavirus diarrhea.