应用多位点串联重复序列分析方法鉴定布鲁氏菌变异株

田国忠; 姜海; 赵鸿雁; 杨晓雯; 朴东日

doi:10.3784/j.issn.1003-9961.2020.07.020

应用多位点串联重复序列分析方法鉴定布鲁氏菌变异株

Identification of Brucella mutants by multiple locus variable-number tandem repeat analysis assay

摘要

摘要:
目的对常规的生物学鉴定方法无法准确分类的布鲁氏菌变异菌，采用多位点串联重复序列分析（MLVA）方法进行种型分类。
方法 39株经常规生物学方法鉴定为布鲁氏菌变异株，应用MLVA方法进行种型分类。
结果 39株菌株与粗糙型血清R凝集，可被Bk₂噬菌体裂解，Wb、Tb噬菌体不裂解，其生物型别不确定；9株菌株与血清A和M弱凝集，2株菌株与血清M弱凝集，其他28株菌株与血清A和M都不凝集；33株菌株可被粗糙型Iz噬菌体裂解，确定39株菌株为布鲁氏菌变异株。应用16个位点串联重复序列分析，结合布鲁氏菌参考标准菌株，经聚类分析，39株菌中2株菌与牛种布鲁氏菌生物2和4型高度相似，37菌株与羊种布鲁氏菌生物3型高度相似。
结论常规生物学鉴定方法与分子生物学分型方法的联合运用，可以更好地解决布鲁氏菌分离菌株的分类，尤其是适合布鲁氏菌变异菌株的鉴定。

Abstract:
ObjectiveTo classify Brucella variants by using multiple locus variable-number tandem repeat analysis (MLVA) assay.
MethodsThirty nine clinical Brucella isolates were identified as Brucella variants by biological methods. They were classified again by MLVA assay.
ResultsThirty nine strains Brucella isolates agglutinated with rough type serum R and could be also cleaved by BK2 phage. But Wb and Tb phages were not cleaved. The biological types were uncertain. 9 strains could agglutinate weakly with serum A and M. 2 strains could agglutinate weakly with serum M. The other 28 strains did not agglutinate with serum A and M. The 39 strains were identified as Brucella variants by biological detection methods. The MLVA assay including 16 tandem repeats loci were used to analyze the 39 Brucella isolates and 17 Brucella reference strains (6 species and types). Cluster analysis showed that in the 39 strains of Brucella variants, 2 strains were similar to Brucella abortus biovar 2 or 4, and 37 strains were similar to Brucella melitensis biovar 3.
ConclusionThe conventional biological identification methods combined with MLVA assay seems to be suitable for further classification of Brucella strains, especially for Brucella variant.

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