| 张鑫,张文竹,李文革,赵红庆,武艳华,李虎,刘正洁,吴媛,卢金星.我国艰难梭菌核糖体分型库的标准化及应用[J].中华流行病学杂志,2019,40(12):1624-1628 |
| 我国艰难梭菌核糖体分型库的标准化及应用 |
| Standardization and application on ribotyping library of Clostridioides difficile in China |
| 收稿日期:2019-05-06 出版日期:2019-12-20 |
| DOI：10.3760/cma.j.issn.0254-6450.2019.12.022 |
| 中文关键词: 艰难梭菌 毛细管电泳 核糖体分型库 |
| 英文关键词: Clostridioides difficile Capillary electrophoresis Ribotyping library |
| 基金项目:国家科技重大专项（2018ZX10733-402） |
| 作者 | 单位 | E-mail | | 张鑫 | 河北北方学院医学检验学院2015级1班, 张家口 075000
中国疾病预防控制中心传染病预防控制所, 北京 102206 | | | 张文竹 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | | | 李文革 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | | | 赵红庆 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | | | 武艳华 | 河北北方学院医学检验学院预防医学教研室, 张家口 075000 | | | 李虎 | 中国疾病预防控制中心传染病预防控制所, 北京 102206
新疆维吾尔自治区阿克苏地区疾病预防控制中心 843000 | | | 刘正洁 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | | | 吴媛 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | wuyuan@icdc.cn | | 卢金星 | 中国疾病预防控制中心传染病预防控制所, 北京 102206 | |
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| 中文摘要: |
| 目的 建立中国艰难梭菌核糖体分型的标准操作流程并补充和验证核糖体分型数据库，为提高各级网络实验室数据间可比性和建立我国艰难梭菌监测网络提供解决方案。方法 参照国外和本实验室已有的核糖体分型操作流程制定实验室标准核糖体分型操作程序，建立基于BioNumerics 7.6软件分析的数据库使用方法，应用此分型程序和数据库使用方法分析54株欧洲地区及美国标准菌株来评价该数据库，对我国2010-2018年13个城市收集的374株艰难梭菌分离菌株进行分型以补充数据库信息，采用Kappa一致性检验方法分析验证实验一致性强弱。结果 欧洲地区及美国标准菌株的毛细管电泳结果条带清晰、位置稳定，聚类结果快速准确；补充分型结果显示，此次分离株中共出现84种型别，其中25种RT型别与国外标准菌株型别一致，而其余的58种型别不同，并且在40种国外标准菌株型别中，有15种型别本次分型未出现；两次验证结果一致性检验Kappa值为0.891（P<0.01），两次测定结果具有一致性，其一致性强度为极强。结论 应用基于QIAxcel毛细管电泳仪建立的标准实验室分型操作流程，所获得的电泳结果具有条带清晰、位置稳定的特点，标准化后的核糖体分型库具有良好的实验室间数据可比性和数据共享便捷性。 |
| 英文摘要: |
| Objective To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China. Methods Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency. Results Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance. Conclusions The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories. |
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