WEI Yu-mei,HE Xiao-qing,CHENG Gang,et al.Primers Selection and Its Application of Detecting theEnterococcus esp Gene based on PCR Method[J].HEILONGJIANG AGRICULTURAL SCIENCES,2011,(04):6-10.
基于PCR技术检测肠球菌esp基因的引物选择及其初步应用
- Title:
- Primers Selection and Its Application of Detecting theEnterococcus esp Gene based on PCR Method
- 文章编号:
- 1002-2767（2011）04-0006-04
- Keywords:
- water environment; MST; enterococcus; esp gene; PCR detection
- 分类号:
- R515
- 文献标志码:
- A
- 摘要:
- 现今全球水域环境受到人类活动的影响，粪便污染日趋严重，因此确定水环境是否存在粪便污染对公众健康、环境治理等方面具有重要意义。以人类肠道致病病原体肠球菌中的esp基因作为检测水体中人类粪便污染的分子标记，从3对具有代表性的引物中选择最佳引物，建立PCR检测方法，得到了较佳的PCR反应条件，并初步应用于湖北武汉地区南湖水中的肠球菌检测。结果表明：利用引物对2可扩增出esp基因片段，其最佳退火温度为52℃。同时在南湖水样中能够检测到阳性结果，证明了引物对2扩增esp基因的可靠性。
- Abstract:
- Nowadays the global water environment has been seriously threatened by human activities，and the fecal contamination become increasingly serious.Therefore to ascertain whether the water environment suffer from fecal pollution is significant for public health and environmental management etc.In this paper，esp gene located in the pathogenic bacteria， enterococcus，in human enteric canal was introduced to work as a molecular marker for detecting human fecal pollution in water environment.And three pairs of typical primers were discussed in PCR method.Then the optimum primers and PCR reaction conditions were obtained and were applied to detect Nanhu water samples in Wuhan，Hubei province.The result indicated that the sequence of esp gene could be amplified by the optimum primers，and its optimum annealing temperature was 52℃.Meanwhile，the water samples in Nanhu which showed the positive results in enterococcus detection verified the reliability of the optimum primers.
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备注/Memo
基金项目：国家水体污染控制与治理科技重大专项资助项目（2009ZX07527-005）；国家“863”资助项目（2008AA062501），中国科学院生态环境研究中心环境水质学国家重点实验室开放基金资助项目（2009-001）