| 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].Chinese journal of Epidemiology,2008,29(5):473-477 |
| 猪链球菌脉冲场凝胶电泳分析方法的建立 |
| Development of a protocol on pulsed neld gel electrophoresis analysis for Streptococcus suits |
| Received:January 21, 2008 |
| DOI： |
| KeyWord: 猪链球菌 脉冲场凝胶电泳分析 |
| English Key Word: Streptococcus suits Pulsed field gel electrophoresis analysis |
| FundProject:国家自然科学基金资助项目（30470095）；科技部“973”计划资助项目（2005cB522904） |
| Author Name | Affiliation | E-mail | | WANG Li-li | 北京市疾病预防控制中心 100013 | | | YE Chang-yun | 北京市疾病预防控制中心 100013 | yechangyun@icdc.cn | | XU Yan-mei | 北京市疾病预防控制中心 100013 | | | CUI Zhi-gang | 北京市疾病预防控制中心 100013 | | | JING Huai-qi | 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 | | | JIN Dong | 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 | | | DU Hua-mao | 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 | | | ZHANG Shou-yin | 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 | | | BAI Xue-mei | 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 | | | ZHAO Ai-lan | 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 | | | XU Jian-guo | 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: |
| 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。 |
| English 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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