CDV-N重组狂犬病病毒的毒力及其免疫原性测定
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摘要
为了探究CDV-N重组狂犬病病毒的毒力及对小鼠的免疫效果,将CDV-N重组狂犬病病毒于BSR细胞上扩毒培养、浓缩后测定其FFD_(50)及LD_(50);将浓缩后的重组病毒液与复合佐剂按一定比例混匀,按不同免疫剂量、不同免疫方式分组免疫接种小鼠,利用ELISA试剂盒对狂犬病、犬瘟热的特异性抗体进行定性/定量检测。结果显示,重组病毒浓缩后的FFD_(50)值为7.85logFFD_(50)/0.1 mL,LD_(50)值为7.67logLD_(50)/0.03mL;免疫接种后第14天抗体阳性率达100%,小鼠血清中的特异性抗体IgG水平随着免疫剂量的递增而递增。试验结果可为CDV-N重组狂犬病病毒制备口服弱毒疫苗的进一步研究提供试验数据。
In order to investigate the virulence of CDV-N recombinant rabies virus and its immunological effect on mice,CDV-N recombinant rabies virus was cultured on BSR cells and its FFD_(50)and LD_(50)were determined after concentration.The concentrated recombinant virus solution was mixed with compound adjuvant in a certain proportion.The mice were immunized with different doses and different immunizations.The qualitative and quantitative detections of specific antibodies against rabies and canine distemper using ELISA Kit were conducted.The results showed that the FFD_(50)of concentrated recombinant virus was 7.85logFFD_(50)/0.1mL,LD_(50)was 7.67logLD_(50)/0.03mL.On the 14th day after immunization,the positive rate of antibody was 100%,and the level of specific antibody IgG in serum of mice increased with the increase of immune dose.The results provided experimental data for further study on preparation of oral attenuated rabies virus vaccine with CDV-N recombinant rabies virus.
In order to investigate the virulence of CDV-N recombinant rabies virus and its immunological effect on mice,CDV-N recombinant rabies virus was cultured on BSR cells and its FFD_(50)and LD_(50)were determined after concentration.The concentrated recombinant virus solution was mixed with compound adjuvant in a certain proportion.The mice were immunized with different doses and different immunizations.The qualitative and quantitative detections of specific antibodies against rabies and canine distemper using ELISA Kit were conducted.The results showed that the FFD_(50)of concentrated recombinant virus was 7.85logFFD_(50)/0.1mL,LD_(50)was 7.67logLD_(50)/0.03mL.On the 14th day after immunization,the positive rate of antibody was 100%,and the level of specific antibody IgG in serum of mice increased with the increase of immune dose.The results provided experimental data for further study on preparation of oral attenuated rabies virus vaccine with CDV-N recombinant rabies virus.
引文
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[2]Oswald M,Geissler S,Goepferich A.Targeting the central nervous system(CNS):a review of rabies virus-targeting strategies[J].Mol Pharm,2017,14(7):2177-2196.
[3]张旭,纪森林,赵雯,等.狂犬病病毒致病机制的最新研究进展[J].中国兽医科学,2018,48(3):295-304.
[4]Fisher C R,Streicker D G,Schnell M J.The spread and evolution of rabies virus:conquering new frontiers[J].Nat Rev Microbiol,2018,16(4):241.
[5]Loots A K,Mitchell E,Dalton D L,et al.Advances in canine distemper virus pathogenesis research:a wildlife perspective[J].J Gen Virol,2017,98(3):311-321.
[6]Yamada K,Ito N,Takayama-Ito M,et al.Multigenic relation to the attenuation of rabies virus[J].Microbiol Immunol,2006,50(1):25-32.
[7]谢世宏.狂犬病[M].陕西西安:陕西科学技术出版社,2005.
[8]Faber M,Pulmanausahakul R,Hodawadekar S S,et al.Overexpression of the rabies virus glycoprotein results in enhancement of apoptosis and antiviral immune response[J].J Virol,2002,76(7):3374-3381.
[9]Faber M,Li J,Kean R B,et al.Effective preexposure and postexposure prophylaxis of rabies with a highly attenuated recombinant rabies virus[J].Proc Natl Acad Sci U S A,2009,106(27):11300-11305.
[10]王喜军.表达狂犬病病毒G蛋白重组犬瘟热弱毒疫苗株的研究[D].北京:中国农业科学院,2012.
[11]刘澜,李士成,陈小云,等.重组表达3G蛋白狂犬病病毒株的构建及重组病毒相关特性的研究[J].中国兽医科学,2016,46(3):326-331.
[12]冯冉,何朝,王凤双,等.北京市顺义区狂犬病风险评估及防控策略[J].中国媒介生物学及控制杂志,2018,29(1):83-87.
[13]周航,李昱,陈瑞丰,等.狂犬病预防控制技术指南(2016版)[J].中华流行病学杂志,2016,37(2):139-163.
[14]赵静.表达犬瘟热病毒H基因重组狂犬病病毒生物学特性的研究[D].广东广州:华南农业大学,2016.
[15]da Fontoura Budaszewski R,Hudacek A,Sawatsky B,et al.Inactivated recombinant rabies viruses displaying canine distemper virus glycoproteins induce protective immunity against both pathogens[J].J Virol,2017,91(8).pii:e02077-16.doi:10.1128/JVI.02077-16.