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Article: Optimization of culture conditions for culturing of influenza virus H1N1 in MDCK cells and its purification
| Title | Optimization 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 | |
| Keywords | Influenza virus MDCK MOI Hemagglutination (HA) TCID50 |
| Issue Date | 2012 |
| Publisher | Zhongguo 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 Identifier | http://hdl.handle.net/10722/191470 |
| ISSN |
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.author | Feng, T | en_US |
| dc.contributor.author | Yin, Z | en_US |
| dc.contributor.author | Li, J | en_US |
| dc.contributor.author | Fan, Y | en_US |
| dc.contributor.author | Tu, W | en_US |
| dc.contributor.author | Li, H | - |
| dc.date.accessioned | 2013-10-15T07:01:27Z | - |
| dc.date.available | 2013-10-15T07:01:27Z | - |
| dc.date.issued | 2012 | en_US |
| dc.identifier.citation | 中國病原生物學雜誌, 2012, v. 7 n. 7, p. 493-496 | en_US |
| dc.identifier.citation | Journal of Pathogen Biology, 2012, v. 7 n. 7, p. 493-496 | - |
| dc.identifier.issn | 1673-5234 | - |
| dc.identifier.uri | http://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.language | chi | en_US |
| dc.publisher | Zhongguo 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.ispartof | Journal of Pathogen Biology | - |
| dc.subject | Influenza virus | - |
| dc.subject | MDCK | - |
| dc.subject | MOI | - |
| dc.subject | Hemagglutination (HA) | - |
| dc.subject | TCID50 | - |
| dc.title | Optimization of culture conditions for culturing of influenza virus H1N1 in MDCK cells and its purification | en_US |
| dc.title | 流感病毒H1N1在MDCK細胞的培養及純化條件的優化 | - |
| dc.type | Article | en_US |
| dc.identifier.email | Tu, W: wwtu@hku.hk | en_US |
| dc.identifier.authority | Tu, W=rp00416 | en_US |
| dc.identifier.hkuros | 225617 | en_US |
| dc.identifier.volume | 7 | - |
| dc.identifier.issue | 7 | - |
| dc.identifier.spage | 493 | - |
| dc.identifier.epage | 496 | - |
| dc.publisher.place | China (中國) | - |
| dc.identifier.issnl | 1673-5234 | - |