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目的 培育 3个不同地理株柔嫩艾美球虫 (Eimeriatenella)的杂交株 ,以探讨研制球虫疫苗的可能性。 方法 通过免疫试验 ,从 5个不同地理株中选择 3株作为杂交亲本株 ,对此 3株分别进行两次杂交 ,获得的后代混合卵囊经单卵囊分离、扩增 ,分别提取卵囊DNA ,利用随机扩增多态性DNA(RAPD)进行分析 ( 3 0条引物 ) ,分离、培育出两代杂交株。再进行免疫试验 ,比较杂交株与亲本株的免疫保护性。 结果 选择了免疫原性较好、免疫保护率较高的广州株、保定株、长春株作为杂交亲本 ,分别进行保定株×长春株、广州株×F1株两次杂交 ,获得的后代卵囊 ,提取卵囊DNA ,RAPD分析后 ,得到了保定株×长春株的杂交株F1(F1Z7)和广州株×F1株的杂交株F2 (F2Z3 )。免疫试验结果 ,杂交株F1与F2的免疫保护率分别为 80 %和 84% ;亲本株广州株为 77% ,保定株为 69% ,长春株为 63 %。 结论 分离、培育出了F1、F2两代杂交虫株。其免疫保护率均高于各亲本株 ,提示杂交株获得了亲本株的部分保护性 ,其免疫的雏鸡对各虫株的攻击均有好的保护力。尤其是F2株 ,免疫保护率平均达到 84%
Objective To develop three hybrid strains of Eimeriatenella with different geographic strains to explore the possibility of developing a coccidiosis vaccine. Methods Three strains from 5 different geographic strains were selected as hybrid parental strains by immunological tests. The three hybrids were crossed twice, respectively. The progeny mixed oocysts were separated and expanded by single oocysts, and oocyst DNA was extracted separately , Using random amplified polymorphic DNA (RAPD) analysis (30 primers), isolated and bred two generations of hybrid strains. Further immune tests were performed to compare the immunoprotective properties of the hybrid and parental strains. Results Guangzhou, Baoding and Changchun strains with better immunogenicity and higher immunoprotection rate were selected as hybrid parents. Two progeny, oocysts, The oocyst DNA was extracted and analyzed by RAPD analysis. F1 (F1Z7) and F2Z3 (F2Z3) of Guangzhou × F1 strain were obtained. The result of immunoassay showed that the immunoprotection rates of hybrid F1 and F2 were 80% and 84% respectively. The parent strain Guangzhou 77%, Baoding 69% and Changchun 63%. Conclusion Isolation, breeding F1, F2 two generations of hybrid strains. The immune protection rate was higher than the parental strains, suggesting that the hybrid strains obtained partial protection of the parental strain, the immune of the chickens on each strain of attack have good protection. Especially F2 strain, the average rate of immune protection reached 84%