Hps Antibodies Established Microtiter Plate Agglutination Test Methods And Research Trivalent Inactivated Vaccine

Haemophilus parasuis belong Pasteurellaceae families genus Haemophilus, it is one of the porcine respiratory commensal bacteria, That can invade the body under certain conditions, Serous fibrinous pleurisy, meningitis, pericarditis and arthritis is characterized by systemic bacterial diseases of pig. H. parasuis morbidity and mortality showed a significant upward trend,it has become Pig industry one of the world’s most severely affected bacterial diseases. pig industry also caused heavy economic losses of China’s. The main results this experiment as follows:1. In the pathogenesis of pig farms to collect suspected diseased samples Haemophilus of16provinces and cities and isolated H. parasuis168. By colony morphology, aspects of cell morphology, staining characteristics and biochemical reactions are in line with the basic characteristics of H. parasuis, PCR analysis also confirmed the isolates indeed H. parasuis, Showed that the prevalence of H. parasuis popular in farm country.2. The H. parasuis isolates were serotype by KRG and ERIC-PCR genotyping parting, Which results KRG serotype4type36(21.43%),5type31(18.45%),13type32(19.05%), the total proportion of58.93%, Accounting virulent serotype (virulent virulent serotype and middle) proportion76.74%. ERIC-PCR genotyping results of ERIC-PCR fingerprinting and standard strains of serotype4ERIC-PCR fingerprinting same proportion of14.29%, accounting for11.31%5-type,13type accounted for13.69%,52.98%of the total number. Comprehensive two typing methods showed H. parasuis serotype4,5type,13type strains are prevalent serotypes of the pig industry, it is Pig pathogenic or potentially pathogenic serotypes main.3. Research on the biological characteristics of isolates showed PT091118h～20h number of viable cells cultured in liquid medium peaked TSB, H.parasuis grown in TSB liquid medium, pH values from7.4to6.2, indicating that the bacteria produce acid strong ability. And the number of viable cells for H.parasuis OD600of statistical analysis, Both showed a linear correlation y=15.244x-0.1959, the return value of R2=0.9727. Based on H.parasuis the number of viable cells and the relationship between the OD600and the number of viable time to reach Summit, Will be able to accurately grasp H.parasuis best time for the preparation of bacterium Haemophilus vaccine and the content value of viable bacteria.4. Antibody microtiter plate agglutination test methods by establishing parasuis, Optimization of reaction Conditions and sensitivity of the optimal working concentration of antigen measured 5.0×109CFu/ml, And have good specificity and reproducibility.5. For the period2011to2013, serum antibody levels in serum samples pigs testing of18provinces and cities in china, the experiment statistical analysis show that Total pig serum samples7414, the positive3567copies,3847copies of negative total positive rate48.11%. Yunnan positive rate of56.02%, Shanghai positive rate57.66%, while the positive rate was19.29%in Guizhou; Nursery pigs (35d-70d) parasuis infection probability reached60.49%, while piglets (0-2Id), sows and boars parasuis infection probability of only about2%; The positive rate was46.91%in spring, The positive rate was was44.03%in Summer,and the positive rate was50.15%in autumn, winter, the positive rate was51.70%. In conclusion, the different regions (between provinces), different growth stages pig (piglet stage, nursery stage, fattening stage, sows, boars) and in different seasons (spring, summer, autumn, winter) the difference between the prevalence of both.6. Preparation of mineral oil adjuvant, aluminum plastic,206and GEL adjuvant trivalent inactivated vaccine Haemophilus parasuis which Screened for preparing a vaccine serotypes4,5type,13type virulent strain. Use of mice, rabbits and pigs as experimental animals to Evaluate its safet. After evaluation of safety, Different adjuvant inactivated vaccine were injected guinea pigs, measuring antibody titers were28-、210、26、211. GEL adjuvant and glue aluminum adjuvant relatively high of antibody titers. The pigs injected antibody titer stable antibody titers decreased situation does not appear. After weighing compares, The aluminum is more suitable for the preparation of gel adjuvant parasuis trivalent inactivated vaccine adjuvant. Alhydrogel trivalent inactivated H.parasuis vaccine protective efficacy experiment piglets, Experimental results piglets incidence of20%, more than the overall protection rate of90%, Showed that alhydrogel vaccinated piglets after the invasion of H.parasuis strains with good resilience. And maternal antibody Experimental results passive protection of piglets, the aluminum plastic inactivated vaccination of pregnant sows producing piglets21days when his protection rate of80%, Showed that inactivated vaccine inoculation The aluminum plastic protective immunity in piglets can delay time, If used the same type of vaccine injection at this stage to stimulate protective immunity in piglets access piglet nursery stage can resist the invasion of H. parasuis.