| 猪链球菌脉冲场凝胶电泳分析方法的建立 |
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| 引用本文： | 王丽丽,叶长芸,许彦梅,崔志刚,景怀琦,金东,杜华茂,张守印,白雪梅,赵爱兰,徐建国.猪链球菌脉冲场凝胶电泳分析方法的建立[J].中华流行病学杂志,2008,29(5):473-477. |
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| 作者姓名： | 王丽丽 叶长芸 许彦梅 崔志刚 景怀琦 金东 杜华茂 张守印 白雪梅 赵爱兰 徐建国 |
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| 作者单位： | 1. 北京市疾病预防控制中心,100013
2. 中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室,北京,102206 |
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| 基金项目： | 国家自然科学基金，国家重点基础研究发展计划(973计划) |
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| 摘 要： | 目的 建立一种可共享的猪链球菌脉冲场凝胶电泳分析(PFGE)方法.方法 利用PulseNet技术优化猪链球菌PFGE分子分型方法,提出新的分析方案,包括胶块的制备、内切酶的选择、电泳条件等.利用DNAStar的Mapdraw软件对猪链球菌基因组序列进行分析,发现在242种内切酶中,Swa Ⅰ、Sma Ⅰ、Apa Ⅰ种酶比较适合.结果 通过对34个血清型共100株菌的分析,发现使用Swa Ⅰ可获得59种带型,使用Sma Ⅰ可获得53种带型,使用Apa Ⅰ可获得43种带型.其中以Swa Ⅰ的分辨率最好.结论 提出猪链球菌的Swa Ⅰ PFGE分析方法,并对条件进行优化,较原方法所需时间短、图像清晰,更具有可重复性,基本具备推广的条件.
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| 关 键 词： | 猪链球菌 脉冲场凝胶电泳分析 |
| 收稿时间： | 2008/1/21 0:00:00 |
Development of a protocol on pulsed neld gel electrophoresis analysis for Streptococcus suits |
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| WANG Li-li,YE Chang-yun,XU Yan-mei,CUI Zhi-gang,JING Huai-qi,JIN Dong,DU Hua-mao,ZHANG Shou-yin,BAI Xue-mei,ZHAO Ai-lan and XU Jian-guo.Development of a protocol on pulsed neld gel electrophoresis analysis for Streptococcus suits[J].Chinese Journal of Epidemiology,2008,29(5):473-477. |
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| Authors: | WANG Li-li YE Chang-yun XU Yan-mei CUI Zhi-gang JING Huai-qi JIN Dong DU Hua-mao ZHANG Shou-yin BAI Xue-mei ZHAO Ai-lan and XU Jian-guo |
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| Institution: | State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;State Key Laboratory for Infectious Disease Prevention and Control, National Institue for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China |
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| Abstract: | Objective To develop a PFGE protocol for Streptococcus suis.Methods We developed and optimized a PFGE protocol for S.suis,in terms of plug preparation,choice of restriction endonucleases and optimized electrophoresis parameters.By analyzing the genome sequences of S.suis P1/7 with Mapdraw of DNAStar.we found three restriction enzymes,Swa Ⅰ,Sma Ⅰ and Apa Ⅰ,were more suitable than others.Results Analysis of 100 isolates of S.suis including 34 of 35 serotypes identified,59,53 and 43 patterns were obtained from Swa Ⅰ,Sma Ⅰ and Apa Ⅰ restriction,respectively.The enzyme Swa Ⅰ had the greatest power for discrimination ability.Conclusion By optimization of the protocol at various conditions,a rapid,reproducible,economic and practical PFGE method for S.suis was developed. |
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| Keywords: | Streptococcus suis Pulsed field gel electrophoresis analysis |
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