肺炎链球菌脉冲场凝胶电泳分型分析方法的建立与优化  

Establishment and optimization of typing method of pulsed field gel electrophoresis for Streptococcus pneumoniae

作　　者：张永茂[1] 贾肇一[1] 王颖童[1] 何宝花[1] 孙印旗[1] ZHANG Yong-mao;JIA Zhao-yi;WANG Ying-tong;HE Bao-hua;SUN Yin-qi(Hebei Province Center for Disease Control and Prevention,Hebei Key Laboratory of Pathogens and Epidemiology of Infectious Diseases,Shjiazhuang 050021,China)

机构地区：[1]河北省疾病预防控制中心河北省传染病病原鉴定分析与流行病学重点实验室,石家庄050021

出　　处：《医学动物防制》2022年第9期915-918,共4页Journal of Medical Pest Control

基　　金：河北省科技计划项目课题(16277745D);河北省卫生计生委医学科研项目(20170053)。

摘　　要：目的 在河北省传染病病原鉴定分析与流行病学重点实验室建立肺炎链球菌(Streptococcus pneumoniae,Spn)脉冲场凝胶电泳(pulsed field gel electrophoresis, PFGE)分型方法,为河北省肺炎链球菌引起的相关疾病的流行病学监测与调查提供可靠的技术支撑。方法 参照美国PulseNet手册标准方法和《国家致病菌识别网技术手册》,在河北省传染病病原鉴定分析与流行病学重点实验室进行肺炎链球菌PFGE分型实验,同时对缓冲液种类、限制性核酸内切酶浓度、酶切时间和电泳条件进行优化。结果 利用优化后的条件对收集到的肺炎链球菌菌株进行PFGE分型分析,所得到的酶切电泳图谱条带清晰,分辨率高。结论 河北省传染病病原鉴定分析与流行病学重点实验室建立和优化了肺炎链球菌PFGE分型分析方法,今后可用于肺炎链球菌的分子特征分析。Objective To provide reliable technical support of epidemiological surveillance and investigation for related diseases caused by Streptococcus pneumoniae(Spn) in Hebei by setting up a pulsed field gel electrophoresis(PFGE) typing method for pneumococcal in Hebei Key Laboratory for Pathogen Identification and Epidemiology of Infectious Diseases.Methods PFGE parting experiments for Streptococcus pneumoniae were conducted by referring to standard methods provided in the US PulseNet manual and Technical Manual for National Pathogen Identification Network in China, at Hebei Key Laboratory for Pathogen Identification and Epidemiology of Infectious Diseases.The buffer type, concentration of restriction endonuclease, enzyme digestion duration and electrophoresis conditions were optimized in the Laboratory.Results The optimized conditions were used for PFGE typing analysis ON Streptococcus pneumoniae strains, and the band of enzyme digestion electrophoretogram was clear with high resolution.Conclusion The PFGE typing method of Streptococcus pneumoniae is established and optimized in the laboratory, and it is eligible to be used for molecular analysis on Streptococcus pneumoniae in the future.

关 键 词：肺炎链球菌 脉冲场凝胶电泳 酶切电泳图谱 限制性核酸内切酶 缓冲液 溶菌酶 分析 

分 类 号：R378.12[医药卫生—病原生物学]