Ward R L, Knowlton D R, Pierce M J
J Clin Microbiol. 1984 Jun;19(6):748-53. doi: 10.1128/jcm.19.6.748-753.1984.
This study was designed to find methods to reproducibly propagate human rotaviruses from fecal specimens and to determine the relationship between particle numbers and infectivity. Growth of virus was initially compared in primary and continuous lines of monkey kidney cells. Primary cells (African green and cynomolgus monkey kidney) supported virus growth directly from fecal specimens much more efficiently than did continuous lines of African green (CV-1) or rhesus (MA104) monkey kidney cells. Rotaviruses were grown in primary cells from 14 of 14 fecal specimens of different individuals collected over a 3-year period. Although rotaviruses in fecal samples could not always be grown in the continuous cell lines, two passages in primary cells appeared to fully adapt the viruses for propagation in the continuous cell line tested (MA104). The efficiency of rotavirus growth was quantified with five of the fecal isolates. It was calculated that, on the average, 1 out of every 46,000 particles in fecal specimens infected monkey kidney cells. After three passages in primary cells, an average of 1 out of every 6,600 progeny virus particles appeared to be infectious. Thus, rotaviruses in fecal specimens were consistently grown in primary cells, and passage in these cells both increased virus infectivity and adapted the viruses for growth in continuous cell lines.
本研究旨在寻找可重复地从粪便标本中增殖人轮状病毒的方法，并确定病毒颗粒数量与感染性之间的关系。最初在猴肾细胞的原代细胞系和连续传代细胞系中比较病毒的生长情况。原代细胞（非洲绿猴和食蟹猴肾细胞）直接从粪便标本中支持病毒生长的效率远高于非洲绿猴（CV - 1）或恒河猴（MA104）肾细胞的连续传代细胞系。在3年期间收集的来自14个不同个体的14份粪便标本的原代细胞中培养出了轮状病毒。尽管粪便样本中的轮状病毒并非总能在连续细胞系中生长，但在原代细胞中传代两次似乎能使病毒完全适应在测试的连续细胞系（MA104）中增殖。用5株粪便分离株对轮状病毒的生长效率进行了量化。据计算，粪便标本中平均每46,000个颗粒中有1个能感染猴肾细胞。在原代细胞中传代三次后，平均每6,600个子代病毒颗粒中有1个似乎具有感染性。因此，粪便标本中的轮状病毒能在原代细胞中持续生长，在这些细胞中传代既能提高病毒的感染性，又能使病毒适应在连续细胞系中生长。
