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用PCR扩增16SrRNA基因鉴定甲型副伤寒沙门菌细胞壁缺陷突变株

The Identification of Cell Wall Deficient Mutants of Salmonella paratyphi A with 16SrRNA Gene Amplified by PCR

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【作者】 黄劲王和

【Author】 HUANG Jin~1,WANG He~2(1.Department of Biochemistry and Molecular Biology,Guiyang Medical College, Guiyang 550004,China; 2.Department of Microbiology,Guiyang Medical College,Guiyang 550004,China)

【机构】 贵阳医学院生物化学与分子生物学教研室贵阳医学院微生物学教研室 贵州贵阳550004贵州贵阳550004

【摘要】 目的:探讨甲型副伤寒沙门菌细胞壁缺陷突变株的检测与鉴定方法。方法:提取甲型副伤寒沙门菌细胞壁缺陷突变株及其亲代细菌型的染色体DNA,根据甲型副伤寒沙门菌的16SrRNA基因保守序列设计引物,进行PCR扩增,对扩增产物进行琼脂糖凝胶电泳与分析。结果:甲型副伤寒沙门菌细胞壁缺陷突变株16SrRNA基因的PCR扩增产物16SrDNA,在230 bp处出现与其亲代细菌型一致的DNA条带和图谱。结论:甲型副伤寒沙门菌的细胞壁缺陷突变株虽然丧失了常规细菌学方法可检测的绝大多数表型特征,但采用聚合酶链反应方法检测该序列,有利于检测和鉴定不能自发返祖的细胞壁缺陷甲型副伤寒沙门菌。

【Abstract】 Objective: To find a sensitive and effective method for the identification of cell wall deficient mutants(CWDMs) of Salmonella paratyphi A.Methods:Chromosome DNAs of CWDMs and bacterial type of S.paratyphi A were extracted and PCR amplification for 16srRNA genes was conducted with the chromsome DNAs as template and using primers designed after the conservative sequences of 16SrDNA of S.paratyphi A.The products were analyzed by agarose-gel electrophoresis.Results:DNA bands of 16SrRNA PCR products from both CWDMs and the bacterial type of S.paratyphi A were all at 230 bp on the agarose-gel. Conclusions: CWDMs derived from S.paratyphi A have lost most of their phenotypic features which are the foundation for their determination by routine bacteriologic methods.The technique of detecting the 16srRNA gene is more effective and sensitive in identifying CWDMs of S.paratyphi A of which the recognition is difficult with routine bacteriologic methods.

  • 【文献出处】 贵阳医学院学报 ,Journal of Guiyang Medical College , 编辑部邮箱 ,2006年04期
  • 【分类号】R378.22
  • 【被引频次】4
  • 【下载频次】162
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