摘要
草鱼出血病病毒湖南邵阳株(CCHV—873),常规培养条件下能在鱼肾(CIK)细胞上形成直径约2mm的蚀斑。当采用三种缓冲系统(MFM-NaHCO_3、MEM-Tris、MEMHEPES)的培养液在普通大气条件下分别培养CIK细胞时,三天内培养液的pH略有变化,其变化范围在0.2—0.4左右,但细胞生长仍然良好,三者无明显差别。在上述系统,以双相法(培养液-凝胶)进行蚀斑试验时,观察到无机缓冲系统培养液的pH变化较大,有机缓冲系统则较稳定,且蚀斑形成的数量显著不同,后者效价比前者要高出4个数量级。因此,在培养液中加入适量的有机缓冲液代之以CO_2的调节是完全有可能的。
A method for plaquing Grass Carp Hemorrhage Virus-873 has been studied. We tested inorganic(NaHCO_3) and organic(Tris, HEPES) buffer systems in open dish culture and found the combination of organic buffers with bicarbonate to be very promising for both cell growth and viral replication The CIK cells growed well in three media (MEM-NaHCO_3, MEM-Tris and MEMHEPES)under normal atmosphere or 5% CO_2 presure, but the organic buffer alone provided a little good cell growth respose. Two-phase (gel-liquid)media incorporating three buffer systems allowed plaquing either in 5% CO_2 presure or in normal atmosphere, but greater efficiency and sensitity were obtained with the combination of organic buffers and bicarbonate than bicarbonate only, and the former is higher 4 orders of magnitude than the latter in plaque numbers. Therefore, it is possibe for plaque assay with organic buffer with bicarbonate instead of CO_2 incubator in open dish culture.
出处
《病毒学杂志》
CSCD
1990年第3期337-340,共4页
关键词
蚀斑试验
病毒效价
Grass Carp Hemrrhage Virus CIK cell Plaque assay Buffer system
