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Article: Optimization of culture conditions for culturing of influenza virus H1N1 in MDCK cells and its purification

TitleOptimization of culture conditions for culturing of influenza virus H1N1 in MDCK cells and its purification
流感病毒H1N1在MDCK細胞的培養及純化條件的優化
Authors
KeywordsInfluenza virus
MDCK
MOI
Hemagglutination (HA)
TCID50
Issue Date2012
PublisherZhongguo Bing Yuan Sheng Wu Xue Za Zhi Bian Ji Wei Yuan Hui (中國病原生物學雜誌編輯委員會). The Journal's web site is located at http://www.cnki.com.cn/Journal/E-E6-ZISC-2012-07.htm
Citation
中國病原生物學雜誌, 2012, v. 7 n. 7, p. 493-496 How to Cite?
Journal of Pathogen Biology, 2012, v. 7 n. 7, p. 493-496 How to Cite?
Abstract目的优化流感病毒H1N1在狗肾细胞(MDCK)上的培养条件,高效扩增流感病毒。方法对MDCK细胞培养流感病毒时添加的胰酶(TPCK-Trypsin)浓度和培养基的种类进行筛选;比较不同细胞代次的MDCK对流感病毒的易感性;按MOI值0.001、0.01、0.1接种H1N1于MDCK细胞,CPE75%时收获病毒上清,并按MOI值0.001接种H1N1后连续培养,分别于1、2、3、4、5d收获病毒上清,HA试验检测上清病毒滴度,浓缩纯化后行TCID50检测,分别选取最佳接种MOI值和收毒时间;对浓缩纯化病毒的红细胞吸附方法进行检验。结果通过对比,确定TPCK-Trypsin的最佳使用浓度是0.25μg/ml,MEM为培养病毒的最佳培养介质;代次低的MDCK对流感病毒的易感性强于代次高的MDCK细胞;按MOI值0.001接种H1N1于MDCK细胞,第3d收获的上清病毒滴度最高,为1∶1 024;用红细胞吸附法浓缩纯化流感病毒的Log 1/TCID50为8.5。结论选用MEM,添加0.25μg/ml TPCK-Trypsin,使用低代次MDCK,病毒接种含量MOI=0.001为H1N1最佳培养条件,红细胞吸附法能高效浓缩纯化培养上清中的H1N1。
Persistent Identifierhttp://hdl.handle.net/10722/191470
ISSN

 

DC FieldValueLanguage
dc.contributor.authorFeng, Ten_US
dc.contributor.authorYin, Zen_US
dc.contributor.authorLi, Jen_US
dc.contributor.authorFan, Yen_US
dc.contributor.authorTu, Wen_US
dc.contributor.authorLi, H-
dc.date.accessioned2013-10-15T07:01:27Z-
dc.date.available2013-10-15T07:01:27Z-
dc.date.issued2012en_US
dc.identifier.citation中國病原生物學雜誌, 2012, v. 7 n. 7, p. 493-496en_US
dc.identifier.citationJournal of Pathogen Biology, 2012, v. 7 n. 7, p. 493-496-
dc.identifier.issn1673-5234-
dc.identifier.urihttp://hdl.handle.net/10722/191470-
dc.description.abstract目的优化流感病毒H1N1在狗肾细胞(MDCK)上的培养条件,高效扩增流感病毒。方法对MDCK细胞培养流感病毒时添加的胰酶(TPCK-Trypsin)浓度和培养基的种类进行筛选;比较不同细胞代次的MDCK对流感病毒的易感性;按MOI值0.001、0.01、0.1接种H1N1于MDCK细胞,CPE75%时收获病毒上清,并按MOI值0.001接种H1N1后连续培养,分别于1、2、3、4、5d收获病毒上清,HA试验检测上清病毒滴度,浓缩纯化后行TCID50检测,分别选取最佳接种MOI值和收毒时间;对浓缩纯化病毒的红细胞吸附方法进行检验。结果通过对比,确定TPCK-Trypsin的最佳使用浓度是0.25μg/ml,MEM为培养病毒的最佳培养介质;代次低的MDCK对流感病毒的易感性强于代次高的MDCK细胞;按MOI值0.001接种H1N1于MDCK细胞,第3d收获的上清病毒滴度最高,为1∶1 024;用红细胞吸附法浓缩纯化流感病毒的Log 1/TCID50为8.5。结论选用MEM,添加0.25μg/ml TPCK-Trypsin,使用低代次MDCK,病毒接种含量MOI=0.001为H1N1最佳培养条件,红细胞吸附法能高效浓缩纯化培养上清中的H1N1。-
dc.languagechien_US
dc.publisherZhongguo Bing Yuan Sheng Wu Xue Za Zhi Bian Ji Wei Yuan Hui (中國病原生物學雜誌編輯委員會). The Journal's web site is located at http://www.cnki.com.cn/Journal/E-E6-ZISC-2012-07.htm-
dc.relation.ispartof中國病原生物學雜誌en_US
dc.relation.ispartofJournal of Pathogen Biology-
dc.subjectInfluenza virus-
dc.subjectMDCK-
dc.subjectMOI-
dc.subjectHemagglutination (HA)-
dc.subjectTCID50-
dc.titleOptimization of culture conditions for culturing of influenza virus H1N1 in MDCK cells and its purificationen_US
dc.title流感病毒H1N1在MDCK細胞的培養及純化條件的優化-
dc.typeArticleen_US
dc.identifier.emailTu, W: wwtu@hku.hken_US
dc.identifier.authorityTu, W=rp00416en_US
dc.identifier.hkuros225617en_US
dc.identifier.volume7-
dc.identifier.issue7-
dc.identifier.spage493-
dc.identifier.epage496-
dc.publisher.placeChina (中國)-

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