病毒学报

研究甲型肝炎病毒H₂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H₂株感染人二倍体细胞(KMB17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H₂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H₂株快速复制病毒株在KMB17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512～2 048之间，感染性滴度均在8.33 lgCCID₅₀/mL±0.125lgCCID₅₀/mL,H₂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H₂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_2strain of the hepatitis A virus(HAV)in human diploid cells and shortening of the culture period of the HAV. Human diploid cells(KMB17cells)infected with the H_2strain of the HAV were studied upon shortening of the culture time to 10 days and continuous passaging. A library of H_2strains 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_2strain of the HAV was harvested after 10 days of culture in KMB17cells. Continuous passaging from the fifth-to-ninth passage revealed antigen content of 512-2,048 and infectious titer >8.33 lgCCID₅₀/mL± 0.125 lgCCID₅₀/mL. Mature particles of the H_2strain 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_2strain of the HAV:(i)shortened the culture time of the HAV;(ii)was stable;(iii)could be used repeatedly. Hence,the rapid-replicating H_2strain of the HAV has important application value.