中国副溶血弧菌多位点可变数目串联重复序列分型方法的建立

目的建立适合中国分离的副溶血弧菌的多位点可变数目串联重复序列分析（MLVA）方法。方法以我国不同来源的420株副溶血弧菌为研究对象，针对现有的12个可变数目串联重复序列（VNTR）位点使用PCR扩增，产物进行毛细管电泳，对不同位点及其组合的分型能力结果进行分析。结果副溶血弧菌的12个VNTR位点中，VPTR7的扩增率比较低（71.90%），VP1-17不具备多态性（Nei值＝0.00），VP2-07的稳定性差；6个位点的组合（VP1-10、VPTR1、VPTR3、VPTR5、VPTR6、VPTR8）分型方案可将420株副溶血弧菌分为311个MLVA型，并可区分相同的ST型菌株。结论6个位点的MLVA分型方案经济性和区分度最优，建立了适合中国分离副溶血弧菌的MLVA分型方案。

Establishment of multiple-locus variable number tandem repeat analysis assay for Vibrio parahaemolyticus isolated in China

ObjectiveTo establish a multiple-locus variable number tandem repeat analysis (MLVA) assay for Vibrio parahaemolyticus isolated in China.MethodsA total of 420 V. parahaemolyticus strains isolated from different areas in China were used. The 12 variable-number tandem-repeat (VNTR) loci were amplified by PCR and the amplicons were analyzed by capillary electrophoresis. The discriminatory power of different VNTR locus and combinations of VNTR loci were evaluated.ResultsThe amplification rates of the 12 VNTR loci of V. parahaemolyticus were different. Locus VP1-17 showed no polymorphism (Nei value＝0.00), locus VP2-03 showed low polymorphism (Nei value＝0.10) and locus VP2-07 showed highest polymorphism. Locus VPTR7 could only be amplified in 71.90% of the test strains. The 420 V. parahaemolyticus strains could be divided into 311 MLVA types using the 6-locus (VP1-10, VPTR1, VPTR3, VPTR5, VPTR6, and VPTR8) .ConclusionThe established 6-locus MLVA scheme showed satisfactory discriminatory power and economic significance and thus is suitable for the molecular typing of V. parahaemolyticusstrains isolated in China.