脉冲场凝胶电泳用于军团菌分型能力的评价

Evaluation of pulsed-field gel electrophoresis for Legionella subtyping

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 capability of pulsed-field gel electrophoresis（PFGE） for the subtyping of Legionella.Methods A total of 117 Legionella reference strains and environmental strains were analyzed by PFGE with Asc Ⅰ.Combined with the number and distributions of bands in the PFGE patterns,capability of subtyping,distinguishability of patterns and epidemiological information,we evaluated the capability of Asc Ⅰ-PFGE for Legionella subtyping.Results All Legionella pneumophila serotypes strains and 17 non-L.pneumophila strains showed high qualitative PFGE patterns;however,for other 13 non-L.pneumophila serotypes strains,most of the PFGE bands were distributed with small weight.Asc Ⅰ-PFGE could cluster the isolates from each water system into the same or very similar patterns and distinguish the strains with different sources.By Asc Ⅰ-PFGE,L.pneumophila subtyping had a high degree of consistency with epidemiological data of strains.Conclusion Asc Ⅰ-PFGE could be used in the subtyping of L.pneumophila and 17 non-L.pneumophila strains,and be applied in the epidemiological investigation of L.pneumophila disease.