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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 analyze the molecular genetic characteristics of Leptospira spp. Strain producing Leptospira vaccine in China and lay a foundation for the research on molecular genetic quality control method of Leptospira vaccine production strain. Methods: The 16S rRNA gene fragment of Rhizoctonia solani was amplified by PCR and sequenced. The genotyping was performed by using pulse field gel electrophoresis (PFGE) and multilocus sequence analysis (MLST). Results: The sequence alignment of 16S rRNA gene showed that the genotype of the genus Loiselles was virulent. The result of PFGE analysis showed that the whole genome of the vaccine strains varied in size from 100-1 000 kb 11 fragments, which were significantly different from the number, size and distribution characteristics of the fragments of the representative isolates from other serogroups in China. The sequence analysis showed that the sequence of this fragment was 140. Conclusion: The results of molecular genetic information analysis of the strain Rhizoctonia solani Vaccine used in production of leptospira vaccine may provide reference for the subsequent research on the molecular genetic quality control method of strain of Leptospira vaccine.