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目的评价脉冲场凝胶电泳(PFGE)技术用于军团菌属不同种(Legionella species)以及不同血清型菌株的分型能力。方法选用117株10个嗜肺军团菌血清型,30种非嗜肺军团菌的参考菌株和我国环境分离菌株,采用AscⅠ内切酶酶切,进行PFGE。从电泳条带的数量和分布、PFGE的分型力、分辨率和与流行病学资料的相关性等4个方面评价PFGE用于军团菌属不同血清型的嗜肺军团菌以及30个非嗜肺军团菌种分型的能力。结果通过AscⅠ酶切、PFGE电泳后,所有血清型的嗜肺军团菌和17种非嗜肺军团菌能获得条带数量适合、且在电泳图谱上条带分布均匀,而另外13种非嗜肺军团菌株条带集中在小分量片段;PFGE能对所有117株选入实验的军团菌进行分析,分型力为100%;AscⅠ-PFGE将43株不同来源的各血清型嗜肺军团菌分成41个不同的型,分型D值为0.9978,将30株非嗜肺军团各自分为不同的型;AscⅠ-PFGE可以将相同来源的嗜肺军团菌和L.m icdadei军团菌分为相同或者相似的型,成簇聚类。结论 AscⅠ-PFGE对所有血清型的嗜肺军团菌和17种非嗜肺军团菌进行PFGE分型,能够获得高质量的电泳图谱,可成功用于嗜肺军团菌的流行病学调查。
Objective To evaluate the ability of pulsed-field gel electrophoresis (PFGE) to classify Legionella species and strains of different serotypes. Methods A total of 117 strains of Legionella pneumophila serogroups, 30 reference strains of Legionella pneumophila and isolates of environmental isolates from China were selected and digested with AscI endonuclease for PFGE. From the four aspects of the number and distribution of electrophoresis bands, the typing power of PFGE, the resolution and the correlation with epidemiological data, the use of PFGE in Legionella pneumophila with different serotypes of Legionella and 30 non-addicted Legionella species typing ability. Results After AscI digestion and PFGE electrophoresis, all serogroups of Legionella pneumophila and 17 non-Legionella pneumophila strains were able to obtain the appropriate number of bands and the bands were evenly distributed on the electrophoresis pattern while the other 13 non-pulmonary Legionella strains were concentrated in the small fragment; PFGE analysis of all 117 Legionella selected experiment, typing force was 100%; Asc Ⅰ-PFGE 43 different origin of each Legionella pneumophila serogroup was divided into 41 A different type, type D value of 0.9978, the 30 non-legionnaires were divided into different types; Asc I-PFGE Legionella pneumophila Lm icdadei can be divided into the same or similar type Clustering. Conclusion AscⅠ-PFGE genotypes of Legionella pneumophila and 17 non-Legionella pneumophila PFGE genotyping, access to high-quality electrophoresis can be successfully used for Legionella pneumophila epidemiological investigation.