CAJ | 학술논문

将含高滴度小反刍兽疫病毒的新鲜病料研磨后接种VERO DS细胞系进行病毒分离，出现细胞病变后连续传代，取3代以上培养物，进行RT-PCR和间接免疫荧光试验及生长曲线测定。生长曲线测定分别以0.01和0.1 MOI接种VERO DS细胞，每隔12h测定病毒滴度，连续培养168h，绘制病毒生长曲线，分析生长特性。结果显示，病料接种细胞72h后，出现细胞病变，经RT-PCR和间接免疫荧光试验鉴定为小反刍兽疫病毒；生长特性研究显示，0.01和0.1 MOI两种病毒接种量，病毒滴度最高均为106TCID50左右，但峰值出现时间和下降速度存在差异，0.01 MOI接种量出现峰值晚，但是维持高滴度时间较长。本文成功分离一株小反刍兽疫病毒，并对两种接毒量的生长特性差异进行了初步分析，为探索该病毒培养方法和收毒时间提供了借鉴。
장함고적도소반추수역병독적신선병료연마후접충VERO DS세포계진행병독분리，출현세포병변후련속전대，취3대이상배양물，진행RT-PCR화간접면역형광시험급생장곡선측정。생장곡선측정분별이0.01화0.1 MOI접충VERO DS세포，매격12h측정병독적도，련속배양168h，회제병독생장곡선，분석생장특성。결과현시，병료접충세포72h후，출현세포병변，경RT-PCR화간접면역형광시험감정위소반추수역병독；생장특성연구현시，0.01화0.1 MOI량충병독접충량，병독적도최고균위106TCID50좌우，단봉치출현시간화하강속도존재차이，0.01 MOI접충량출현봉치만，단시유지고적도시간교장。본문성공분리일주소반추수역병독，병대량충접독량적생장특성차이진행료초보분석，위탐색해병독배양방법화수독시간제공료차감。
The pathological samples containing high titer of peste des petits ruminants virus(PPRV)were chosen and treated to inoculate VERO DS cell line for PPRV isolation. The third passage cultures were used for PCR test and indirect immuno-fluorescence test to make sure that peste des petits ruminants virus was isolated after CPE appearance in the cell line. In order to understand the virus growth characteristics,VERO DS cell was inoculated with the culture of 0.1 MOI and 0.01 MOI and cultivated for 168 hours and TCID50 was tested every 12 hours. The results showed that CPE appeared 72 hours after inoculation and PPRV was successfully isolated as identified by RT-PCR test and indirect immuno-fluorescence test. The growth curve showed that the highest titer of the two inoculation doses reached nearly 106 TCID50 but with different shapes. The study is hoped to provide references in PPRV isolation and culttivation.