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城市污水中病原性大肠埃希氏菌毒力基因eaeA和rfbE的实时荧光定量PCR检测

谢润欣 张崇淼 王晓昌 孙婷婷 李宪

谢润欣, 张崇淼, 王晓昌, 孙婷婷, 李宪. 城市污水中病原性大肠埃希氏菌毒力基因eaeA和rfbE的实时荧光定量PCR检测[J]. 环境科学研究, 2012, 25(8): 922-926.
引用本文: 谢润欣, 张崇淼, 王晓昌, 孙婷婷, 李宪. 城市污水中病原性大肠埃希氏菌毒力基因eaeA和rfbE的实时荧光定量PCR检测[J]. 环境科学研究, 2012, 25(8): 922-926.
XIE Run-xin, ZHANG Chong-miao, WANG Xiao-chang, SUN Ting-ting, LI Xian. Quantitative Detection of Virulence Genes eaeAand rfbEof Pathogenic Escherichia coliin Municipal Wastewater by Real-Time PCR[J]. Research of Environmental Sciences, 2012, 25(8): 922-926.
Citation: XIE Run-xin, ZHANG Chong-miao, WANG Xiao-chang, SUN Ting-ting, LI Xian. Quantitative Detection of Virulence Genes eaeAand rfbEof Pathogenic Escherichia coliin Municipal Wastewater by Real-Time PCR[J]. Research of Environmental Sciences, 2012, 25(8): 922-926.

城市污水中病原性大肠埃希氏菌毒力基因eaeA和rfbE的实时荧光定量PCR检测

基金项目: 国家自然科学基金项目(50908185);国家环境保护环境微生物利用与安全控制重点实验室开放基金项目(MARC 2011D038);陕西省教育厅专项科研项目(11JK0765)

Quantitative Detection of Virulence Genes eaeAand rfbEof Pathogenic Escherichia coliin Municipal Wastewater by Real-Time PCR

  • 摘要: 针对病原性大肠埃希氏菌EHEC和EPEC的毒力基因eaeA和rfbE,选用特异性引物,建立了实时荧光定量PCR检测方法. 利用从污水中分离出的目的核酸片段构建重组质粒,通过测序和BLAST比对分析,确定了PCR扩增的特异性. 将重组质粒作为模板,分别测定标准曲线,在eaeA基因模板量8.77~8.77×105 copy,rfbE基因模板量4.98~4.98×105copy的范围内,Ct(循环阈值)与模板量的对数值具有良好的线性关系〔R2为0.997(eaeA)和1.000(rfbE)〕. 结合膜吸附洗脱浓缩方法,对于实际水样,eaeA和rfbE基因的检测限分别为1.75×102和9.96×101copy/100 mL. 利用该方法对城市污水处理厂进、出水进行检测的结果显示,污水二级处理工艺对这2种毒力基因的去除效果明显.

     

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出版历程
  • 收稿日期:  2011-11-29
  • 修回日期:  2012-04-19
  • 刊出日期:  2012-08-25

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