高原地区流感病毒培养的条件优化  

Exploration of the Most Suitable Condition for Culturing Influenza Virus on Plateau Region

作　　者：胡澜怀[1] 周霞[1] 

机构地区：[1]攀枝花市疾病预防控制中心

出　　处：《现代检验医学杂志》2016年第3期131-134,共4页Journal of Modern Laboratory Medicine

摘　　要：目的探索高原地区流感病毒在犬肾细胞（MDCK）上的最佳培养条件，提高流感病毒的分离效果。方法将原始标本分别接种于 MDCK细胞，通过比较CO2百分浓度（5％，4.8％，4.6％，4.4％，4.2％），TPCK-胰酶浓度（1，2，3，4，5，6，8，10μg／ml），接种量（100，200，300，400，500μl），培养时间（48，72，96，120，144 h），接种方法（吸附接种，直接接种），培养容器（细胞瓶，6孔板）对流感病毒的影响，确定最佳培养条件，对分离的各型病毒用红细胞凝集（HA）检测病毒滴度。结果通过病毒血凝素滴度结果比较，4.4％ CO2与4.6％ CO2，2μg／ml TPCK-胰酶，300μl与400μl的接种量、吸附接种、72 h～96 h的培养时间、6孔板收获的病毒血凝素滴度高。结论在高原地区实验室建立了 MDCK细胞培养流感病毒的最优条件。Objective To explore the most suitable condition of influenza virus which was culturing on plateau region,and im-prove the effect of the separation of influenza virus.Methods The original specimens were respective inoculated in MDCK cells,by comparing CO2 percent concentration （5%,4.8%,4.6%,4.4%,4.2%）,TPCK-pancreatic enzyme dosage （1,2,3, 4,5μg/m and 6,8,10μg/ml）,inoculation amount （100,200,300,400,500μl）,incubation time （48,72,96,120,144 h）,inoc-ulation methods （adsorption,direct inoculated）,and culture vessel（cells,6 orifice）that were influenced on influenza virus, the best culture conditions was determined.Hemagglutinatio （HA）method was used to detect the virus titers.Results Through the comparison,the HA titer of virus was in the highest titer with 6 orifice plate culture vessel,4.4% CO2 and 4.6% CO2 ,2μg/ml TPCK-pancreatic enzyme,300μl and 400μl inoculation amount,72h^96h of incubation time.Conclusion The optimal condition of the influenza virus cultured with the MDCK cell has been established in the laboratory on plateau region.

关 键 词：流感病毒 犬肾细胞 培养条件 

分 类 号：R373.13[医药卫生—病原生物学] R446.5[医药卫生—基础医学]