柯萨奇病毒A组10型高效价免疫血清的制备和应用分析  

Preparation and application of immune sera with high-titre anti-coxsackievirus A10 polyclonal antiboby

作　　者：卫星辰 谢天宏[1] 李华[1] 唐东红[1] 岳磊[1] 叶尤松[1] 张宇昊 赵红 向虹 王杰 杨婷[1] WEI Xing-chen;XIE Tian-hong;LI Hua;TANG Dong-hong;YUE Lei;YE You-song;ZHANG Yu-hao;ZHAO Hong;XIANG Hong;WANG Jie;YANG Ting(Institute of Medical Biology,Chinese Acaclemy of Medical Sciences,Kunming,Yunman 650118,China)

机构地区：[1]中国医学科学院,北京协和医学院医学生物学研究所,云南昆明650118

出　　处：《中国病毒病杂志》2022年第1期21-25,共5页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金项目(32000654);云南省科技计划项目基础研究专项(202001AT070147);云南省医学后备人才培养计划项目(H-2019065);中国医学科学院医学与健康科技创新工程(2021-1-I2M-043)。

摘　　要：目的利用CVA10不同毒株制备高效价免疫血清,并探索血清制备的影响因素和应用效果。方法将CVA10-2044株和CVA10-N25株皮下免疫家兔、山羊和大鼠,或腹腔注射大鼠,测定血清中和抗体效价,分析不同抗原类型、动物种属及免疫途径对抗体应答的影响;用免疫血清对系列稀释的病毒进行棋盘中和,以确定完全中和不同剂量病毒所需血清量;中和法评价该免疫血清应用于病毒株鉴定、外源因子检测的效果。结果 CVA10-2044株纯化液经4次免疫后家兔中和抗体效价可达1∶4 096;CVA10-N25株收获液腹腔或浓缩液皮下和肌内免疫大鼠,2次或3次免疫后效价最高达1∶4 096和1∶6 144;N25株纯化液免疫山羊和家兔,4次免疫后效价最高可达1∶4 096。研究发现,完全中和50 000 CCID;病毒所需血清量为32 U,病毒量低于5 CCID_(50)时,1 U血清即可完全中和。2个毒株制备的免疫血清应用于病毒株鉴定结果和RT-PCR法鉴别结果一致。结论 CVA10-2044株或CVA10-N25株纯化液皮下免疫家兔或山羊、N25株收获液腹腔免疫大鼠均可获得高效价免疫血清,该血清可应用于病毒株鉴定和外源因子检测。Objective To prepare immune sera with high-titre anti-coxsackievirus A10(CVA10)polyclonalantibody and to explore the impact factors for the preparation and the application of immune sera.Methods The rabbits,goats or rats were immunized with CVA10-2044 and CVA10-N25 strain,then the neutralizing antibody titre of sera against CVA10 were determined.The impact factors for the antibody response to different antigens in different animals through various immune routes were discussed.A series of diluted CVA10 viruses were neutralized with the prepared immune serum to determine the required minimum amount of anti-CVA10 serum,and application of the immune serum was further assessed by the identification of virus isolates.Results The neutralizing antibody titer in rabbits immunized four times with purified CVA10-2044 strain was 1∶4096,and the highest titers in rats challenged with two or three doses of CAV10-N25 strain subcutaneously or intramuscularly were 1∶4096 and 1∶6144,respectively,and the highest titer in rabbits or goats after four doses of purified N25 strain was 1∶4096.It was found that 32 U of anti-CVA10 immune serum was sufficient to neutralize 50000 CCID_(50)of viruses,while 1 U of serum was able to fully neutralize virus of<5 CCID;.It was further demonstrated that identification of virus isolates using the immune serum prepared by both 2044 strain and N25 strain was consistent with the RT-PCR method.Conclusions Immune sera with high-titre of anti-CVA10 polyclonal antibody can be obtained in rabbits or goats subcutaueously immunized with purified CAV10-2044 or CAV10-N25 viruses»or in rats intraperitoneally inoculated with CAV10-N25 virus,which may be used for the identification of virus isolates and detection of exogenous factors.

关 键 词：免疫血清 柯萨奇病毒A组10型 中和抗体 血清制备 

分 类 号：R373.23[医药卫生—病原生物学]