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首页> 外文期刊>Environmental Science & Technology >Real Time PCR for the Rapid Detection of vanA Gene in Surface Waters and Aquatic Macrophyte by Molecular Beacon Probe
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Real Time PCR for the Rapid Detection of vanA Gene in Surface Waters and Aquatic Macrophyte by Molecular Beacon Probe

机译：利用分子信标探针实时PCR快速检测地表水和水生植物中的vanA基因

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摘要

Enterococci serve as an "indicator" of fecal contamination for recreational water quality. The vancomycin-resistant-enterococci (VRE) are emerging environmental contaminants in the surface waters. The aim of this study was to develop a rapid and specific molecular beacon probe (MBP)-based real-time PCR assay for detection of vanA gene in surface waters and aquatic macrophyte. The limit of detection (LOD) of the MBP assay was 1 CFU/mL of VRE [r = 0.943; PCR efficiency = 99.7%] in 2-fold dilution format within 2.5 h and demonstrated high specificity for environmental enterococci isolates exhibiting VanA phenotype (n = 25). VRE were detected from downstream surface waters of the rivers impacted by point sources of pollution and recreational activities. The probe detected vanA gene in root-mat associated microbiota of E crassipes (Mart.) Solms. an aquatic nuisance weed, at eutrophic sites of the surface waters (ANOVA p < 0.001). In addition, the assay enabled detection of otherwise nondetectable vanA gene concentration in the upstream sites of two Indian rivers (Student's t test p < 0.001). The MBP assay developed can be used for sensitive and rapid detection of VRE in surface waters and identification of nonpoint sources of pollution for implementation of preventive measures to protect human health.
机译：肠球菌是休闲水质粪便污染的“指标”。耐万古霉素肠球菌（VRE）是地表水中新兴的环境污染物。这项研究的目的是开发一种基于快速和特异性分子信标探针（MBP）的实时PCR检测试剂盒，用于检测地表水和水生植物中的vanA基因。 MBP测定的检出限（LOD）为1 CFU / mL的VRE [r = 0.943;在2.5小时内以2倍稀释形式的PCR效率= 99.7％]，并显示出对表现出VanA表型的环境肠球菌分离物具有高度特异性（n = 25）。从受污染和娱乐活动的主要来源影响的河流下游地表水中检测到VRE。探针在E. crassipes（Mart。）Solms的根-垫相关微生物群中检测到vanA基因。在地表水的富营养化位置的水生杂草（ANOVA p <0.001）。此外，该测定能够检测两条印度河流上游站点中原本无法检测到的vanA基因浓度（Student's t检验p <0.001）。开发的MBP测定法可用于灵敏和快速检测地表水中的VRE，并识别非点源污染，以实施预防措施来保护人类健康。

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  • 来源
    《Environmental Science & Technology》 |2009年第9期|3343-3348|共6页
  • 作者

    PUSHPA LATA; SIYA RAM; MADHOOLIKA AGRAWAL; RISHI SHANKER;

  • 作者单位

    Indian Institute of Toxicology Research (CSIR), P.O. Box 80, Mahatma Gandhi Marg, Lucknow-226001, U.P., India;

    Indian Institute of Toxicology Research (CSIR), P.O. Box 80, Mahatma Gandhi Marg, Lucknow-226001, U.P., India;

    Banaras Hindu University, Varanasi-221005, U.P., India;

    Indian Institute of Toxicology Research (CSIR), P.O. Box 80, Mahatma Gandhi Marg, Lucknow-226001, U.P., India;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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