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目的采用多位点可变数目串联重复序列分析技术(MLVA)分析贵州省黑线姬鼠钩端螺旋体(简称钩体)分离株的分子流行病学特征,为贵州省钩体病的防控提供依据。方法采用硅胶膜型TM细菌基因组DNA提取试剂盒提取56株致病性钩体DNA,选择7个可变数目串联重复序列VNTR位点进行PCR扩增,扩增产物经2%琼脂糖凝胶电泳后用凝胶成像系统成像并计算各VNTR位点的扩增长度,分析菌株各VNTR位点的重复数目;采用BioNumerics软件分析,用UPGMA系数分析分离株与参考菌株间的聚类关系。结果 MLVA分析结果显示,56株钩端螺旋体有VNTR4、VNTR36、VNTR50位点出现重复数目差别,被分为6个MLVA型(A1、B1、B2、B3、B4、B5型)。聚类分析显示56株钩端螺旋体与参考菌株黄疸出血群56601株聚类关系较近,与其他参考株聚类关系较远。结论贵州省近年黑线姬鼠钩端螺旋体分离株均为黄疸出血群,MLVA型别具有多样性。
Objective To analyze the molecular epidemiological characteristics of Leptospira interrogans in Guizhou Apodemus agrarius by MLVA (Multiple Site Variable Number Tandem Repeat Analysis) in accordance with. Methods 56 strains of pathogenicity leptospira DNA were extracted by using genomic DNA extraction kit of genomic DNA from genomic DNA. Seven VNTR sites with variable number of tandem repeats were selected for PCR amplification. The amplified products were separated by 2% agarose gel electrophoresis The length of each VNTR locus was calculated by gel imaging system and the number of VNTR loci was analyzed. BioNumerics software was used to analyze the clustering relationship between isolates and reference strains by UPGMA coefficient. Results The results of MLVA analysis showed that there were 6 MLVA types (A1, B1, B2, B3, B4 and B5) in 56 leptospira strains with VNTR4, VNTR36 and VNTR50 sites. Cluster analysis showed that 56 strains of Leptospira were closely related to 56601 strains of reference strains of jaundice hemorrhage and clustering with other reference strains. CONCLUSION: Leptospira interrogans of Apodemus agrarius in recent years in Guizhou Province are both hemorrhagic jaundice and MLVA genotypes are diverse.