Schmidt N J, Ho H H, Riggs J L, Lennette E H
Appl Environ Microbiol. 1978 Sep;36(3):480-6. doi: 10.1128/aem.36.3.480-486.1978.
In efforts to define the most sensitive cell culture systems for recovery of viruses from wastewaters, 181 samples were inoculated in parallel into tube cultures of various cell types and were plaqued in bottle and petri dish cultures of three types of monkey kidney cells. Polioviruses were recovered most frequently in the RD line of human rhabdomyosarcoma cells, group A coxsackieviruses in RD and human fetal diploid kidney (HFDK) cells, group B coxsackieviruses in the BGM line of African green monkey kidney cells, echoviruses in RD and primary rhesus monkey kidney (RhMK) cells, and reoviruses in RhMK cells. BGM cells were unsatisfactory for recovery of viruses other than polioviruses and group B coxsackieviruses, and a line of fetal rhesus monkey kidney (MFK) was not a satisfactory substitute for primary RhMK. With RhMK cells, comparable numbers of virus isolations were made in tube cultures and in plaque assays conducted in bottle cultures, but with BGM and MFK cells, fewer isolations were made by plaquing than by inoculation of tube cultures. In comparative plaque assays on fecal samples under three different overlays in bottle and plate cultures of RhMK, BGM, and MFK cells, it was found that plaquing in the most sensitive system, RhMK, was less efficient for virus recovery than was inoculation of tube cultures of RhMK or HFDK cells. Overall, plaque assays performed in petri dishes in a CO(2) incubator yielded fewer virus isolates than did parallel plaque assays performed in closed bottle cultures. Other limitations of plaque assays for recovery of human enteric viruses are discussed.
为了确定从废水中分离病毒最敏感的细胞培养系统,将181份样品同时接种到各种细胞类型的试管培养物中,并在三种猴肾细胞的瓶式和平皿培养物中进行空斑测定。在人横纹肌肉瘤细胞的RD系中最常分离到脊髓灰质炎病毒,在RD和人胚二倍体肾(HFDK)细胞中最常分离到A组柯萨奇病毒,在非洲绿猴肾细胞的BGM系中最常分离到B组柯萨奇病毒,在RD和恒河猴原代肾(RhMK)细胞中最常分离到埃可病毒,在RhMK细胞中最常分离到呼肠孤病毒。BGM细胞对于分离脊髓灰质炎病毒和B组柯萨奇病毒以外的病毒并不理想,而恒河猴胎儿肾(MFK)细胞系也不能很好地替代原代RhMK细胞。对于RhMK细胞,在试管培养物和瓶式培养物中的空斑测定中分离到的病毒数量相当,但对于BGM和MFK细胞,空斑测定分离到的病毒比试管培养接种法少。在RhMK、BGM和MFK细胞的瓶式和平皿培养物中,对粪便样品在三种不同覆盖物下进行比较空斑测定,发现最敏感的系统RhMK中的空斑测定在病毒分离效率上低于RhMK或HFDK细胞的试管培养接种法。总体而言,在CO₂培养箱中的平皿中进行的空斑测定比在封闭瓶式培养中进行的平行空斑测定分离到的病毒更少。还讨论了空斑测定在分离人肠道病毒方面的其他局限性。