TPCK胰酶和普通胰酶对各亚型流感病毒在MDCK细胞分离培养中的影响  

Effect of TPCK-trypsin and trypsin on the MDCK cells culture and isolation of subtype influenza virus

作　　者：马萍[1] 李欣玥 MA Ping LI Xin-yue(Viral Laboratory, Shaanxi Center for Disease Control and Prevention, Xi＇an, Shaanxi 714000, China)

机构地区：[1]陕西省疾病预防控制中心病毒室,陕西西安710054 [2]渭南市疾病预防控制中心,陕西渭南714000

出　　处：《中国卫生检验杂志》2017年第18期2585-2587,2592,共4页Chinese Journal of Health Laboratory Technology

基　　金："十二五"国家科技重大专项(2013ZX10004202-001-002)

摘　　要：目的比较不同种类的胰酶对不同亚型流感病毒在MDCK细胞增殖能力的影响,寻找最佳胰酶类型和浓度。方法将血凝滴度为1∶16的甲型H1N1、H3N2、Victoria、Yamagata流感病毒稀释100倍接种到MDCK细胞上,分别改变维持液中TPCK-胰酶和普通胰酶浓度,在24 h、48 h、72 h、96 h取培养物上清测血凝滴度,比较不同亚型流感病毒在MDCK细胞病变速度和病毒含量的差异。结果 TPCK胰酶对各亚型流感病毒在MDCK细胞培养中的增值能力明显提高,TPCK胰酶在甲型H1N1、H3N2、Victoria、Yamagata流感病毒的最优浓度分别为1μg/ml^3μg/ml、1μg/ml、0.5μg/ml^2μg/ml、1μg/ml。普通胰酶对甲型H1N1流感病毒的增值能力有提高作用,最佳胰酶浓度为5μg/ml,普通胰酶对H3N2、Victoria、Yamagata流感病毒在MDCK细胞的增值无提高作用。结论 MDCK培养流感病毒时,需要添加TPCK胰酶。Objective To compare the effects of different types of trypsin on the proliferation of different subtypes of influenza virus culture on MDCK cells,so as to find the best trypsin type and concentration. Methods The influenza virus H1N1,H3N2,Victoria and Yamagata with hemagglutination titer of 1∶16 were diluted 100 times and cultivated on MDCK cell in different culture conditions,the different concentrations of TPCK trypsin and common trypsin were changed,and then we observed CPE and measured hemagglutination titer at 24 h,48 h,72 h and 96 h. The differences in speed and virus content among different subtypes of influenza viruses in MDCK cells were compared. Results TPCK significantly increased the multiplication of influenza subtypes on MDCK cell culture. The optimal concentration of TPCK trypsin in A（ H1N1）,H3N2,Victoria,Yamagata influenza virus was within 1 μg/ml-3 μg/ml,1 μg/ml,0. 5 μg/ml-2 μg/ml and 1 μg/ml. Normal trypsin can increase the A（ H1N1） influenza multiplication on MDCK cells. The optimal concentration of trypsin in H1N1 was 5 μg/ml. Nomal ttrypsin can not increase the multiplication of H3N2,Victoria and Yamagata influenza on MDCK cells. Conclusion When we culture influenza virus on MDCK,we need to add TPCK-trypsin.

关 键 词：流感 细胞培养 胰酶 

分 类 号：R511.7[医药卫生—内科学]