中国钩端螺旋体疫苗生产用菌种罗株分子遗传特性分析

Molecular genetic analysis of strain Luo for production of Chinese Leptospira vaccine

目的：分析研究中国钩端螺旋体疫苗生产用菌种罗株的分子遗传特性,为钩体疫苗生产用菌种分子遗传质量控制方法的研究奠定基础。方法：通过对罗株16S rRNA基因片段进行PCR扩增、测序,并运用脉冲场凝胶电泳（PFGE）和多位点序列分析（MLST）等方法,对其进行分子分型。结果：16S rRNA基因序列比对结果显示钩体疫苗生产用菌种罗株基因种为致病性问号型;PFGE分析结果表明该疫苗株全基因组在酶切后共有100～1 000 kb大小不等的片段11个,与我国其他不同血清群代表株酶切片段的数量、大小和分布特征方面明显不同;多位点序列分析得出该罗株序列型为140。结论：本文对钩体疫苗生产用菌种罗株的分子遗传信息分析结果,可为后续的钩体疫苗生产用菌种分子遗传质量控制方法的研究提供参考。

Objective： To understand the molecular genetic background of strain Luo for production of Leptospira vaccine in China,and provide the foundation for the genetic quality control method of strains for production of Leptospira vaccine in the future. Methods： PCR method was used to amplify 16 S rRNA gene and PCR product was sent for sequencing. Pulsed field gel electrophoresis（ PFGE） and multiple locus sequence typing（ MLST） were used to analyze types of the vaccine strain Luo. Results： The alignment analysis of 16 S rRNA gene sequencing revealed that the genetic species of strain Luo for vaccine production was pathogenic Leptospira interrogans. It was observed that the PFGE profile of strain Luo consisted of 11 gene fragments of 100-1 000 kb,which was different from the number,size and distribution characteristics of the representative strains of other different serogroups in China. MLST showed that the ST type of strain Luo was 140. Conclusion： The molecular genetic information of strain Luo for production of Leptospira vaccine obtained in this study provides the reference for the research on genetic quality control methods of strains for Leptospira vaccine production in the future.