利用狗肾传代细胞分离流感病毒及分离条件的研究

发布时间：2018-01-12 03:10

  本文关键词：利用狗肾传代细胞分离流感病毒及分离条件的研究　出处：《实用医学杂志》2014年05期 　论文类型：期刊论文

  更多相关文章： 流感 人 MDCK细胞 流感病毒 分离 谷氨酰胺

【摘要】：目的:对首次利用狗肾传代细胞(MDCK)分离流感病毒进行总结,摸索流感病毒细胞培养分离技术,积累分离的经验,提高流感监测水平。方法:培养MDCK细胞,并接种61例经咸阳市疾病预防控制中心流感网络实验室PCR检测为阳性的标本,收集病变细胞,经红细胞凝集实验、红细胞凝集抑制实验进行流感病毒毒株鉴定,同时在培养液中加入不同浓度谷氨酰胺以研究谷氨酰胺对MDCK细胞生长的影响。结果:从61例实时荧光定量PCR检测阳性流感病例标本中分离出25株流感毒株,HA滴度均≥1∶16,血凝抑制实验(HI)结果显示HI效价均≥1∶640,其中18株为A(H3N2)亚型株,7株为A(H1N1)亚型株;谷氨酰胺浓度降至0.1%以下时,细胞生长速率低,细胞培养至48 h活性即开始下降。结论:利用MDCK细胞培养分离流感病毒在咸阳市取得成功,并发现谷氨酰胺浓度对MDCK细胞的活性和细胞凋亡有着显著的影响。
[Abstract]:Objective: to summarize the method of isolation of influenza virus by using MDCK for the first time, to explore the technique of cell culture and isolation of influenza virus, and to accumulate the experience of isolation. Methods: MDCK cells were cultured and 61 samples were inoculated with positive PCR from influenza network laboratory of the Center for Disease Control and Prevention of Xianyang City to collect pathological cells. The influenza virus strains were identified by erythrocyte agglutination test and erythrocyte agglutination inhibition test. At the same time, different concentrations of glutamine were added to the culture medium to study the effect of glutamine on the growth of MDCK cells. Twenty-five influenza strains were isolated from 61 cases of influenza positive by real-time fluorescence quantitative PCR. HA titer 鈮，

本文编号：1412421