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首页> 外文期刊>Food microbiology >Development of a real-time PCR assay with an internal amplification control for detection of Gram-negative histamine-producing bacteria in fish
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Development of a real-time PCR assay with an internal amplification control for detection of Gram-negative histamine-producing bacteria in fish

机译：带有内部扩增控制的实时PCR分析方法的开发，用于检测鱼中革兰氏阴性的组胺产生细菌

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摘要

Prompt detection of bacteria that contribute to scombrotoxin (histamine) fish poisoning can aid in the detection of potentially toxic fish products and prevent the occurrence of illness. We report development of the first real-time PCR method for rapid detection of Gram-negative histamine-producing bacteria (HPB) in fish. The real-time PCR assay was 100% inclusive for detecting high-histamine producing isolates and did not detect any of the low- or non-histamine producing isolates. The efficiency of the assay with/ without internal amplification control ranged from 96-104% and in the presence of background flora and inhibitory matrices was 92/100% and 73-96%, respectively. This assay was used to detect HPB from naturally contaminated yellowfin tuna, bluefish, and false albacore samples. Photobacterium damselae (8), Plesiomonas shigelloides (2), Shewanella sp. (1), and Morganella morganii (1) were subsequently isolated from the real-time PCR positive fish samples. These results indicate that the real-time PCR assay developed in this study is a rapid and sensitive method for detecting high-HPB. The assay may be adapted for quantification of HPB, either directly or with an MPN-PCR method.
机译：及时发现会导致中毒（组胺）鱼中毒的细菌，有助于发现潜在有毒的鱼产品并预防疾病的发生。我们报告了第一个实时PCR方法的发展，用于快速检测鱼中革兰氏阴性组胺生产细菌（HPB）。实时PCR检测包括100％的检测高组胺产生分离株的检测，没有检测任何低或非组胺生产分离株的检测。具有/不具有内部扩增对照的测定的效率范围为96-104％，并且在背景菌群和抑制性基质存在的情况下分别为92/100％和73-96％。该测定法用于检测自然污染的黄鳍金枪鱼，蓝鱼和假长鳍金枪鱼样品中的HPB。 damselae细菌（8），志贺菌Plesiomonas shigelloides（2），Shewanella sp。 （1）和摩根氏摩根氏菌（1）随后从实时PCR阳性鱼样品中分离出来。这些结果表明，在这项研究中开发的实时PCR分析是一种快速，灵敏的检测高HPB的方法。该测定可以直接或通过MPN-PCR方法适于HPB的定量。

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  • 来源
    《Food microbiology》 |2011年第3期|p.356-363|共8页
  • 作者

    Kristin Bjornsdottir-Butler; Jessica L. Jones; Ronald Benner; William Burkhardt III;

  • 作者单位

    Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, 1 Ibervilte Dr., Dauphin Island, AL 36528, USA;

    Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, 1 Ibervilte Dr., Dauphin Island, AL 36528, USA;

    Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, 1 Ibervilte Dr., Dauphin Island, AL 36528, USA;

    Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, 1 Ibervilte Dr., Dauphin Island, AL 36528, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    histamine; biogenic amines; histidine decarboxylase; real-time PCR; bacteria; fish;

    机译：组胺;生物胺组氨酸脱羧酶;实时PCR;菌;鱼;

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