本文關(guān)鍵詞： 流感病毒 MDCK細(xì)胞 分離培養(yǎng)　出處：《現(xiàn)代預(yù)防醫(yī)學(xué)》2017年23期 　論文類型：期刊論文

【摘要】：目的優(yōu)化不同亞型流感病毒在狗腎上皮細(xì)胞(MDCK)上最優(yōu)培養(yǎng)條件,提高不同亞型流感病毒在MDCK細(xì)胞上的分離效果。方法將甲型H1N1、H3N2、Victoria、Yamagata四種亞型的流感病毒以不同的培養(yǎng)條件:不同培養(yǎng)溫度、不同培養(yǎng)基、不同的牛血清濃度、不同L-1(對(duì)-甲苯磺酰)苯丙胺酰氯甲酮(TPCK)胰酶濃度、不同的接種量、不同的接種時(shí)間接種到MDCK細(xì)胞上,在24h、48h、72h、96h取培養(yǎng)物上清測(cè)血凝滴度,比較不同亞型流感病毒在MDCK細(xì)胞病變速度和病毒含量的差異。結(jié)果甲型H1N1流感病毒的最優(yōu)培養(yǎng)溫度為35℃,DMEM培養(yǎng)基,無牛血清,TPCK胰酶濃度為1-3μg/ml,接種300ml,不吸附或吸附1.5-3.0小時(shí);H3N2病毒的最優(yōu)培養(yǎng)溫度為34℃,DMEM培養(yǎng)基,無牛血清,TPCK胰酶濃度為1μg/ml,吸附1.5小時(shí),接種300ml;Victoria流感病毒的最優(yōu)培養(yǎng)溫度為35℃,DMEM培養(yǎng)基,無牛血清,TPCK胰酶濃度為0.5-2.0μg/ml,接種200ml,吸附1.5小時(shí);Yamagata流感病毒的最優(yōu)培養(yǎng)溫度為35℃,DMEM培養(yǎng)基,無牛血清,TPCK胰酶濃度為1μg/ml,接種200ml,吸附1.5小時(shí)。結(jié)論根據(jù)流感病毒不同亞型來選擇流感病毒的培養(yǎng)條件可獲得更好的實(shí)驗(yàn)結(jié)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，

本文編號(hào)：1461259