摘要
目的应用布鲁氏菌种多重PCR和猪种多重PCR对布鲁氏菌进行鉴定。方法对两套多重PCR方法进行条件优化,应用布鲁氏菌参考菌株、疫苗株和临床分离菌株进行评价,并与生物学分型方法进行比较。结果布鲁氏菌种多重PCR扩增(包括牛种布鲁氏菌8个生物型,猪种5个生物型,羊种3个生物型,以及绵羊附睾、犬和沙漠森林野鼠种各1个生物型)可获得152～1 682bp的8个DNA片段,但种间DNA片段数不一。猪种多重PCR扩增猪种5个生物型菌株得到197～774bp的7个DNA片段,不同生物型的DNA扩增图谱不同;猪种疫苗株S2扩增图谱与猪种生物1型相同,多重PCR分型方法与生物学分型方法分析结果一致。结论布鲁氏菌种多重PCR能鉴别布鲁氏菌6个种,猪种多重PCR可以鉴别猪种5个生物型。两种多重PCR均具有较好的特异性和重复性,该方法快速、简便,可推广应用。
Objective To evaluate the ability of two types of multiplex PCR assays （Bruce ladder and Suis-ladder） to distinguish six Brucella species and five biovars of Brucelia suis. Methods A Bruce-ladder multiplex PCR assay was performed using 8 pairs of primers to distinguish six Brucella species. A Suis ladder multiplex PCR assay was performed using 4 pairs of primers to distinguish five biovars of B. suis. The tested strains, which consisted of 27 reference strains of six Brucella species and 239 Brucella isolates, were examined using PCR assays and methods of biological identifica-tion. Results The Bruce-ladder multiplex PCR assay differentiated six Brucella species, including Brucella abortus, Brucella rnelitensis , Brucella suis , Brucella canis , Brucella ovis , and Brucella neotornae , in a single step. The Suis-lad-der multiplex PCR assay differentiated five biovars of B. suis in a single step. The electrophoresis patterns of the vaccine strain S2 and biovar 1 of B. suis were identical. The total rate of concordance between methods of biological identification and multiplex PCR assays was 100%. Conclusion The Bruce-ladder and Suis-ladder multiplex PCR assays are rapid and specific methods of respectively identifying Bruceila species and B. suis at the biovar level.
出处
《中国病原生物学杂志》
CSCD
北大核心
2013年第2期126-130,共5页
Journal of Pathogen Biology
关键词
布鲁氏菌
菌种
生物型
多重PCR
鉴别
Brucella
species
biovar
multiplex PCR assays
identification
