狂犬病毒在Vero细胞上的传代适应

本文研究了3株狂犬病毒(C_(42)、C_(103)及4aG株)在Vero细胞上的培养条件和方法,并通过连续传代培养,选出1株滴度平均达到7.13LogLD_(50)/ml以上,达到WHO规定的不需浓缩的标准(≥10~(7.0)ICLD_(50)/ml)毒株—C_(42)—V_(170)病毒的增殖高峰由原来的10天左右提前到3～5天。另外2株的病毒滴度也达到了WHO规定的制备人用Vero细胞狂犬疫苗(PVRV)的标准(≥10~(6.0)ICLD_(50)/ml)。3株病毒都能在Vero细胞上产生蚀斑,符合WHO提出的疫苗生产毒种的要求。

Adaptation and passages of rabies virus strains in vero cell cultures

Rabies is a lethal widespread epidemic. At present,only means of prevention and cure of rabies is injection of rabies vaccine.WHO has recently recommended the Purified Vero Rabies Vaccine(PVRV). Vero cells were chosen as a substrate for rabies vaccine production because of the cell growth potential in microcarrier culture and the high virus yield,i.e, conditions suitable for an economical large-scale production of inactivated rabies vaccine.Potency and stability of PVRV have been satisfactory, equivalent to or higher than human diploid cell vaccine.The study and preparation of PVRV have just begun in our country.The important factor in the production of vaccine is to increase the titer of virus, which was studied and explored in this paper.The culture condition and passage methods of CTN-1-42, CTN-1-103, 4aG three rabies virus strains were studied in vero cells. The titer of virus suspension in each passage were determined by mice(i. e)test and plaque formation test after 8 or 9 passages. The results showed that:(l)The mean titer of C42-Vl7 strain was about 7.13 LogLD 50/ml through 17 passages in vero cells, which reached the standard of unnecessary concentration of virus harvest bv WHO regulation. (2) The incubation period in vero cells was shortened from about 10 days to 3～5 days. The titer of other two virus strains(≥10~6.5LogLD_(50)/ml)reachcd the standard for the preparation of PVRV(≥10~6.0Log LD_(50)/ml).