牛病毒性腹泻病毒在MDBK细胞上培养增殖影响因素的研究  

Study on the factors influencing the proliferation of Bovine viral diarrhea virus in MDBK cells

作　　者：魏玲[1] 梁洪 王怀禹[1] 吕远蓉[1] 杨国淋 WEI Ling;LIANG Hong;WANG Huaiyu;LYU Yuanrong;YANG Guolin(Nanchong Vocational and Technical College,Nanchong 637131,China;North Sichuan Medical College,Nanchong 637100,China)

机构地区：[1]南充职业技术学院,四川南充637131 [2]川北医学院,四川南充637100

出　　处：《黑龙江畜牧兽医》2021年第17期66-70,145,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：南充市科技计划项目(19YFZJ0009)。

摘　　要：为了确定牛病毒性腹泻病毒(Bovine viral diarrhea virus, BVDV)在牛肾细胞(Madin darby bovine kidey cell, MDBK细胞)上的培养增殖影响因素,试验通过优化接毒剂量、吸附时间、收毒时间、血清浓度、培养液pH值等参数,确定BVDV在MDBK细胞上增殖的最佳培养条件,并利用该培养条件增殖培养BVDV及进行免疫原性试验,研究其对犊牛的免疫原性。结果表明:BVDV在MDBK细胞上培养的最佳接毒量为毒种稀释1×10^(3)倍,最佳吸附时间为1 h,最佳收毒时间为接毒后48小时,最佳血清浓度为2%,最佳培养液pH值为7.2,应用最佳培养条件培养的3批次培养物的病毒含量分别为1.0×10^(6.88)TCID_(50)/0.1 mL、1.0×10^(6.83)TCID_(50)/0.1 mL、1.0×10^(6.88)TCID_(50)/0.1 mL,利用该培养物制备的油乳剂灭活疫苗在免疫犊牛后第7天即可产生BVDV中和抗体,至免疫后第180天中和抗体效价仍维持在较高水平,具有良好的免疫原性。说明通过优化培养条件确定了BVDV在MDBK细胞上的增殖培养影响因素,利用该培养条件增殖培养的BVDV的病毒液具有良好的免疫原性。In order to determine the factors influencing the proliferation of Bovine viral diarrhea virus(BVDV) in Madin darby bovine kidey cells(MDBK cells),the experiment determined the influencing factors of BVDV culture and proliferation on MDBK cells by optimizing the changes of the parameters such as different inoculation amounts, different adsorption time, different collection time, different serum concentrations, and different pH values of culture fluid;BVDV was cultivated by using the determined proliferation influencing factors for the study of its immunogenicity on calves. The results showed that the optimal dose of BVDV cultured on MDBK cells was 1×10^(3)-fold dilution of the virus seed, the best adsorption time being 1 h, the best collection time being 48 h after inoculation, and the best serum concentration being 2%,the optimal pH value of the culture medium being 7.2. The virus contents of the three batches of culture under the best culture conditions were 1.0×10^(6.88)TCID_(50)/0.1 mL,1.0×10^(6.83)TCID_(50)/0.1 mL,1.0×10^(6.88)TCID_(50)/0.1 mL;the inactivated vaccine prepared from the culture produced BVDV neutralizing antibodies seven days after immunization of the calves, and the neutralizing antibodies remained at a high level 180 days after immunization;and it had good immunogenicity. The results suggested that through the optimization of the culture conditions, the factors influencing the culture and proliferation of BVDV on MDBK cells have been grasped;the BVDV virus liquid that proliferated under this culture condition had good immunogenicity.

关 键 词：牛病毒性腹泻病毒 MDBK细胞 培养 增殖 影响因素 病毒含量 中和抗体 免疫原性 

分 类 号：S852.653[农业科学—基础兽医学]