流感病毒在狗肾上皮细胞上的最优培养条件

发布时间：2018-01-24 22:57

  本文关键词： 流感病毒 MDCK细胞 分离培养　出处：《现代预防医学》2017年23期 　论文类型：期刊论文

【摘要】：目的优化不同亚型流感病毒在狗肾上皮细胞(MDCK)上最优培养条件,提高不同亚型流感病毒在MDCK细胞上的分离效果。方法将甲型H1N1、H3N2、Victoria、Yamagata四种亚型的流感病毒以不同的培养条件:不同培养温度、不同培养基、不同的牛血清浓度、不同L-1(对-甲苯磺酰)苯丙胺酰氯甲酮(TPCK)胰酶浓度、不同的接种量、不同的接种时间接种到MDCK细胞上,在24h、48h、72h、96h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。结果甲型H1N1流感病毒的最优培养温度为35℃,DMEM培养基,无牛血清,TPCK胰酶浓度为1-3μg/ml,接种300ml,不吸附或吸附1.5-3.0小时;H3N2病毒的最优培养温度为34℃,DMEM培养基,无牛血清,TPCK胰酶浓度为1μg/ml,吸附1.5小时,接种300ml;Victoria流感病毒的最优培养温度为35℃,DMEM培养基,无牛血清,TPCK胰酶浓度为0.5-2.0μg/ml,接种200ml,吸附1.5小时;Yamagata流感病毒的最优培养温度为35℃,DMEM培养基,无牛血清,TPCK胰酶浓度为1μg/ml,接种200ml,吸附1.5小时。结论根据流感病毒不同亚型来选择流感病毒的培养条件可获得更好的实验结果。
[Abstract]:Objective to optimize the optimal culture conditions of different subtypes of influenza virus on dog renal epithelial cells (MDCK) and to improve the isolation of different subtypes of influenza virus from MDCK cells. Four subtypes of influenza virus H3N2VictoriaVictoria yagata were cultured under different culture conditions: different culture temperature, different culture medium and different bovine serum concentration. Different L-1 (p-toluenesulfonyl) phenylalanine acetyl ketone (TPCK) trypsin concentration, different inoculation amount and different inoculation time were inoculated on MDCK cells for 72 h at 24 h. Hemagglutination titer of the supernatant was measured at 96 h to compare the pathological changes of different subtypes of influenza virus in MDCK cells and the virus content. Results the optimal culture temperature of influenza A (H1N1) virus was 35 鈩，

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