进口轮状病毒疫苗RotaTeq的热稳定性  

Thermal stability of imported rotavirus vaccine RotaTeq

作　　者：杜加亮[1] 刘艳[1] 焦洋[2] 于晴川[1] 刘悦越[1] 赵荣荣[1] 高加梅 范行良[1] 国泰[1] DU Jia-liang;LIU Yan;JIAO Yang;YU Qing-chuan;LIU Yue-yue;ZHAO Rong-rong;GAO Jia-mei;FAN Xing-liang;GUO Tai(National Institutes for Food and Drug Control,Beijing 102629,China;不详)

机构地区：[1]中国食品药品检定研究院肠道病毒疫苗室,北京102629 [2]北京市朝阳区疾病预防控制中心微生物检验科,北京100021

出　　处：《中国生物制品学杂志》2020年第8期849-854,共6页Chinese Journal of Biologicals

基　　金：国家科技重大专项课题(2012ZX10004702)。

摘　　要：目的分析进口轮状病毒(rotavirus,RV)疫苗RotaTeq(RV5)的热稳定性。方法采用荧光定量PCR法分别测定22~25和37℃条件下放置不同时间的RV5疫苗滴度,以参考疫苗株滴度为标准,计算RV5疫苗5个型别毒株(G1、G2、G3、G4、P1)滴度和总滴度。同时扩增RV5疫苗G3型毒株的VP7序列,并与GenBank中登录的原型病毒株r G3的VP7序列进行比较。结果22~25℃条件下,G1、G2、G3、G4、P1型毒株滴度和总滴度每天下降速度分别为3.51%、2.26%、3.19%、5.22%、3.18%和3.61%,各型别毒株间差异无统计学意义(P>0.05);37℃条件下,G3型毒株滴度的下降速度(31.20%/h)明显高于其他型别毒株(G1、G2、G4、P1下降速度分别为每小时0.75%、0.31%、1.08%和2.07%),差异有统计学意义(P<0.05)。G3型毒株滴度于37℃的半衰期仅为0.07 d,其原型株rG3的半衰期可达2.10 d。RV5疫苗G3型疫苗株与其原型株的差异包括散在的氨基酸差异(F32L、T69A、L83F、I118T、D151G、P275L)及位于5′UTR(T25C)和3′UTR(G1041A、A1045G、G1046A、G1047A、C1048T、A1053T)较聚集的核苷酸差异。结论RV5疫苗中各型别毒株在22~25℃时热稳定性良好,但在37℃时G3型疫苗株是影响RV5疫苗热稳定性的关键,位于VP7 UTR的核苷酸突变可能是导致其热稳定性降低的主要原因。Objective To analyze the thermal stability of an imported rotavirus vaccine RotaTeq(RV5).Methods The virus titers of RV5 vaccine stored at 22~25℃and 37℃for various time durations were determined by real-time PCR,based on which the titers of vaccine strains of five types(G1,G2,G3,G4 and P1)and the total titer were calculated using the titer of reference vaccine as standard.The VP7 sequence of RV5 vaccine strain of type G3 was amplified and compared with that of prototypic strain r G3 in GenBank.Results The decrease rates of titers of vaccine strains of types G1,G2,G3,G4 and P1 and the total titer were 3.51%,2.26%,3.19%,5.22%,3.18%and 3.61%per day respectively at 25℃,which showed no significant difference between various types(P>0.05).However,at 37℃,the decrease rate of vaccine strain of type G3(31.20%per hour)was significantly higher than those of other types(G1:0.75%;G2:0.31%;G4:1.08%;P1:2.07%)(P<0.05).The half life of titer of vaccine strain of type G3 at 37℃was only 0.07 d,while that of its prototypic strain rG3 was 2.10 d.The RV5 vaccine strain of type G3 was different from its prototypic strain in scattered amino acids(F32 L,T69 A,L83 F,I118 T,D151 G and P275 L)as well as nucleotides in5′untransplated region(UTR)(T25 C)and 3′UTR(G1041 A,A1045 G,G1046 A,G1047 A,C1048 T and A1053 T).Conclusion The RV5 vaccine strains of various types showed high thermal stability at 25℃.However,at 37℃,the low thermal stability of vaccine strain of type G3 showed significant impact on the thermal stability of RV5 vaccine by a possible mechanism of nucleotide mutation in VP7 UTR.

关 键 词：轮状病毒 热稳定性 RotaTeq 非翻译区 

分 类 号：R373.25[医药卫生—病原生物学]