不同灭活条件对山羊副流感病毒3型免疫原性的影响不同灭活条件对山羊副流感病毒3型免疫原性的影响(附件)【字数：5512】
1
目 录
Abstract 1
Key words 2
引言2
1 材料与方法2
1.1 材料2
1.1.1主要试剂2
1.1.2 细胞与病毒2
1.1.3引物2
1.1.4 实验动物2
1.2 方法3
1.2.1 制备病毒抗原3
1.2.2 病毒滴度的测定3
1.2.2.1 1%豚鼠红细胞悬液的制备3
1.2.2.2 血凝试验3
1.2.3病毒的灭活3
1.2.4病毒灭活效果的检测3
1.2.4.1 血凝价检测4
1.2.4.2病毒传代与RTPCR检测 4
1.2.5 小鼠免疫试验4
1.2.6 HI抗体检验4
2结果与分析 4
2.1 灭活后血凝性检测 5
2.2传代与RTPCR检测5
2.3 HI抗体效价检测 8
3讨论 8
致谢8
参考文献9
图1 0.1%甲醛灭活效果的RTPCR检测6
图2 0.3%甲醛灭活效果的RTPCR检测6图3 BPL灭活效果的RTPCR检测6
图4 温度灭活效果的RTPCR检测 6
图5 免疫后HI抗体变化8
表1 动物免疫方案5
表2 甲醛灭活后血凝价变化 5
表3 BPL灭活后血凝价变化 5
表4 温度灭活后血凝价变化 5
表5 一免后各组HI抗体滴度7
表6 二免后各组HI抗体滴度7
不同灭活条件对山羊副流感病毒3型免疫原性的影响
动物药学专业学生 吴铖楠
指导教师 王先炜
摘要：为探索山羊副流感病毒3型(CPIV3)的最佳灭活条件，本实验选择甲醛、β丙内脂（BPL）和温度进行病毒的灭活试验，探索最佳灭活条件，并检测对其免疫原 *好棒文|www.hbsrm.com +Q: *351916072*
性的影响。将山羊副流感病毒3型JS2013株在MDBK细胞上增殖，测得血凝价为128。选择0.1%、0.3%甲醛、0.01%β丙内脂以及56℃水浴分别灭活不同时间，检测病毒血凝价的变化，结果显示仅BPL灭活后对病毒血凝价无显著影响。将灭活的病毒在MDBK细胞传代3代，进行RTPCR检测。结果表明0.1%甲醛12h可将病毒完全灭活，0.3%甲醛4h即可灭活病毒；0.01%的BPL 6h可灭活病毒；56℃作用15min可灭活病毒。选择不同条件灭活病毒，制备油乳剂灭活疫苗。在物理性状、无菌试验检验合格后，经皮下注射免疫ICR小鼠，检测血清HI抗体。结果显示一免后小鼠血清抗体即转阳，二免后达到较高水平。0.1%甲醛灭活效果最佳，温度灭活病毒诱导产生的抗体水平最低。以上结果表明可选择0.1%甲醛作为灭活疫苗制备的灭活剂，这将为CPIV3灭活疫苗的制备提供理论依据。
The effects of different inactivation conditions towards the immunogenicity of Caprine parainfluenza virus type 3(CPIV3)
Student majoring in Veterinary Pharmacy Chengnan Wu
Tutor Xianwei Wang
Abstract：To explore the best inactivation condition for Caprine parainfluenza virus type 3(CPIV3), this?experiment compared?the?inactive effects of?formaldehyde, BPL and temperature, selected the best inactivation condition and tested the effects on immunogenicity of the virus. Firstly, CPIV3 JS2013?strain was proliferated on MDBK cells, with a HA titer of 128 (27). 0.1%, 0.3% formaldehyde, 0.01% BPL and 56℃ waterbath were selected to inactivate virus for different times respectively. The changes of HA titers were tested the results revealed that only BPL had no obvious effect on it. Inactivated viruses were cultured on MDBK for 3 passages and detected by RTPCR. Results indicated that 0.1% formaldehyde inactivated 12h can inactivate virus completely, 0.3%formaldehyde inactivated 4h can inactivate virus; 0.01%BPL inactivated 6h can inactivate virus; 56℃ inactivated 15min can inactivate virus. Oil emulsion?inactivation vaccines were prepared after the virus was inactivated by selected conditions. After the physical characteristics inspection and sterility test, ICR mice were immunized 2 times with 2 weeks interval. Serum samples were collected and tested by HI assay. The results indicated that serum antibody turned positive after the first immunization, and the antibody reached high level after the second immunization. The group of 0.1% formaldehyde showed the best immunogenicity, the level of antibody produced by temperature inactivation was the lowest. These results indicated that 0.1% formaldehydeas could be chosen to prepare CPIV3 inactivation vaccine, this provided theoretical foundation for CPIV3 vaccine study.
目 录
Abstract 1
Key words 2
引言2
1 材料与方法2
1.1 材料2
1.1.1主要试剂2
1.1.2 细胞与病毒2
1.1.3引物2
1.1.4 实验动物2
1.2 方法3
1.2.1 制备病毒抗原3
1.2.2 病毒滴度的测定3
1.2.2.1 1%豚鼠红细胞悬液的制备3
1.2.2.2 血凝试验3
1.2.3病毒的灭活3
1.2.4病毒灭活效果的检测3
1.2.4.1 血凝价检测4
1.2.4.2病毒传代与RTPCR检测 4
1.2.5 小鼠免疫试验4
1.2.6 HI抗体检验4
2结果与分析 4
2.1 灭活后血凝性检测 5
2.2传代与RTPCR检测5
2.3 HI抗体效价检测 8
3讨论 8
致谢8
参考文献9
图1 0.1%甲醛灭活效果的RTPCR检测6
图2 0.3%甲醛灭活效果的RTPCR检测6图3 BPL灭活效果的RTPCR检测6
图4 温度灭活效果的RTPCR检测 6
图5 免疫后HI抗体变化8
表1 动物免疫方案5
表2 甲醛灭活后血凝价变化 5
表3 BPL灭活后血凝价变化 5
表4 温度灭活后血凝价变化 5
表5 一免后各组HI抗体滴度7
表6 二免后各组HI抗体滴度7
不同灭活条件对山羊副流感病毒3型免疫原性的影响
动物药学专业学生 吴铖楠
指导教师 王先炜
摘要：为探索山羊副流感病毒3型(CPIV3)的最佳灭活条件，本实验选择甲醛、β丙内脂（BPL）和温度进行病毒的灭活试验，探索最佳灭活条件，并检测对其免疫原 *好棒文|www.hbsrm.com +Q: *351916072*
性的影响。将山羊副流感病毒3型JS2013株在MDBK细胞上增殖，测得血凝价为128。选择0.1%、0.3%甲醛、0.01%β丙内脂以及56℃水浴分别灭活不同时间，检测病毒血凝价的变化，结果显示仅BPL灭活后对病毒血凝价无显著影响。将灭活的病毒在MDBK细胞传代3代，进行RTPCR检测。结果表明0.1%甲醛12h可将病毒完全灭活，0.3%甲醛4h即可灭活病毒；0.01%的BPL 6h可灭活病毒；56℃作用15min可灭活病毒。选择不同条件灭活病毒，制备油乳剂灭活疫苗。在物理性状、无菌试验检验合格后，经皮下注射免疫ICR小鼠，检测血清HI抗体。结果显示一免后小鼠血清抗体即转阳，二免后达到较高水平。0.1%甲醛灭活效果最佳，温度灭活病毒诱导产生的抗体水平最低。以上结果表明可选择0.1%甲醛作为灭活疫苗制备的灭活剂，这将为CPIV3灭活疫苗的制备提供理论依据。
The effects of different inactivation conditions towards the immunogenicity of Caprine parainfluenza virus type 3(CPIV3)
Student majoring in Veterinary Pharmacy Chengnan Wu
Tutor Xianwei Wang
Abstract：To explore the best inactivation condition for Caprine parainfluenza virus type 3(CPIV3), this?experiment compared?the?inactive effects of?formaldehyde, BPL and temperature, selected the best inactivation condition and tested the effects on immunogenicity of the virus. Firstly, CPIV3 JS2013?strain was proliferated on MDBK cells, with a HA titer of 128 (27). 0.1%, 0.3% formaldehyde, 0.01% BPL and 56℃ waterbath were selected to inactivate virus for different times respectively. The changes of HA titers were tested the results revealed that only BPL had no obvious effect on it. Inactivated viruses were cultured on MDBK for 3 passages and detected by RTPCR. Results indicated that 0.1% formaldehyde inactivated 12h can inactivate virus completely, 0.3%formaldehyde inactivated 4h can inactivate virus; 0.01%BPL inactivated 6h can inactivate virus; 56℃ inactivated 15min can inactivate virus. Oil emulsion?inactivation vaccines were prepared after the virus was inactivated by selected conditions. After the physical characteristics inspection and sterility test, ICR mice were immunized 2 times with 2 weeks interval. Serum samples were collected and tested by HI assay. The results indicated that serum antibody turned positive after the first immunization, and the antibody reached high level after the second immunization. The group of 0.1% formaldehyde showed the best immunogenicity, the level of antibody produced by temperature inactivation was the lowest. These results indicated that 0.1% formaldehydeas could be chosen to prepare CPIV3 inactivation vaccine, this provided theoretical foundation for CPIV3 vaccine study.
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