甲型肝炎病毒H2快速复制株在人二倍体细胞中的增殖研究  

Growth Characteristics of the Rapid-replicating H2Strain of the Hepatitis A Virus in Human Diploid Cells

作　　者：王东宝[1] 马忠飞[1] 刘泽[1] 宋盛波 卢志敏 施红进 何陆涛 李燕[1] 李卫东[1] 廖国阳[1] WANG Dongbao;MA Zhongfei;LIU Ze;SONG Shengbo;LU Zhimin;SHI Hongjin;HE Lutao;LI Yan;LI Weidong;LIAO Guoyang(Institute of Medical Biology Chinese Academy of Medical Sciences,Kunming 650118 China)

机构地区：[1]中国医学科学院医学生物学研究所,昆明650118

出　　处：《病毒学报》2022年第3期627-634,共8页Chinese Journal of Virology

摘　　要：研究甲型肝炎病毒H_(2)快速复制株在人二倍体细胞中的生长特性,并缩短甲肝病毒的培养周期。将甲型肝炎病毒H_(2)株感染人二倍体细胞(KMB_(17)细胞株),采用高病毒感染复数(MOI)将病毒培养时间从26d缩短至10d后收获病毒,并通过连续传代进行适应研究,建立H_(2)株快速复制毒种库,在不同培养时间检测病毒抗原、感染性滴度,绘制病毒生长曲线,进行传代稳定性验证和病毒形态学的观察。甲肝H_(2)株快速复制病毒株在KMB_(17)细胞上培养10d后收获,连续传代从第5代至第9代,抗原含量均在512~2048之间,感染性滴度均在8.33 lgCCID_(50)/mL±0.125lgCCID_(50)/mL,H_(2)株快速繁殖至5代病毒和9代病毒在电镜下观察到多为成熟的实心颗粒。在5批次的重复试验中,病毒培养至10d、16d、22d时,收获的病毒感染性滴度无显著差异(P>0.005)。筛选后的甲型肝炎病毒H_(2)株的快速复制株缩短了甲肝病毒的培养时间,且保持较好的稳定性和重复性,具有重要的应用价值。To study the growth characteristics of the rapid-replicating H_(2) strain of the hepatitis A virus(HAV)in human diploid cells and shortening of the culture period of the HAV.Human diploid cells(KMB_(17)cells)infected with the H_(2) strain of the HAV were studied upon shortening of the culture time to 10 days and continuous passaging.A library of H_(2) strains was constructed.The virus antigen titer and infectious titer were measured at different culture times.A one-step growth curve of the virus was drawn to ascertain the stability of the passage,and we observed viral morphology by electron microscopy.The rapid-replicating H_(2) strain of the HAV was harvested after 10 days of culture in KMB_(17) cells.Continuous passaging from the fifth-to-ninth passage revealed antigen content of 512-2,048 and infectious titer>8.33 lgCCID_(50)/mL±0.125 lgCCID_(50)/mL.Mature particles of the H_(2) strain were observed under electron microscopy.After five batches of replicate testing,a significant difference in the virus titer was not observed until 10,16,and 22 days.The rapid-replicating H_(2) strain of the HAV:(i)shortened the culture time of the HAV;(ii)was stable;(iii)could be used repeatedly.Hence,the rapid-replicating H_(2) strain of the HAV has important application value.

关 键 词：甲肝病毒 H_(2)快速复制株 连续传代 

分 类 号：R373.2[医药卫生—病原生物学]