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首页> 外文学位 >Detection of fish pathogens with PCR-DGGE in non-lethal mucus samples and molecular typing of Aeromonas salmonicida using the 16S-23S internal transcribed spacers.
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Detection of fish pathogens with PCR-DGGE in non-lethal mucus samples and molecular typing of Aeromonas salmonicida using the 16S-23S internal transcribed spacers.

机译：利用PCR-DGGE检测非致死性粘液样本中的鱼类病原体，并使用16S-23S内部转录间隔子对鲑鱼气单胞菌进行分子分型。

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摘要

This thesis is an investigation of the detection of Aeromonas salmonicida using nested PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) in non-lethal fish mucus samples. The technique was compared to conventional non-lethal and lethal methods. A. salmonicida was detected by all techniques. Detection of A. salmonicida by DGGE was based on co-migrating bands and was verified by sequencing. The mean level of detection was highest with culture of mucus on Coomassie brilliant blue agar in the Chinook (71.67% +/- 7.64) and PCR-DGGE (69.10 +/- 1.29) for the coho salmon. Mucus based techniques had a statistically significant agreement for detection of A. salmonicida. Flavobacterium psychrophilum, F. branchiophilun and Yersinia ruckeri were not detected with PCR-DGGE. Bands often co-migrated to these pathogens, but their nucleotide sequences belonged to other bacteria. Seeding Y. ruckeri in mucus showed that PCR-DGGE could detect the pathogen.; In an attempt to detect strain differences, the 16S-23S rRNA internal transcribed spacers of A. salmonicida were fingerprinted using RFLP and DGGE. These techniques verify the clonal nature of the pathogen, but do show the capacity to differentiate various Aeromonas spp.
机译：本文采用嵌套式PCR-变性梯度凝胶电泳（PCR-DGGE）技术检测非致死鱼粘液样品中的鲑鱼气单胞菌。将该技术与常规的非致命和致命方法进行了比较。通过所有技术检测到沙门氏菌。 DGGE对鲑鱼曲霉的检测基于共迁移条带，并通过测序进行了验证。平均检出水平最高的是奇努克族的库马斯亮蓝琼脂上的粘液培养物（71.67％+/- 7.64）和银大麻哈鱼的PCR-DGGE（69.10 +/- 1.29）。基于粘液的技术在检测沙门氏菌中具有统计学意义。 PCR-DGGE未检测到嗜黄杆菌，支链杆菌和rersinia ruckeri。乐队经常共迁移到这些病原体，但它们的核苷酸序列属于其他细菌。在黏液中播种ruckeri.ruckeri表明，PCR-DGGE可以检测病原体。为了检测菌株差异，使用RFLP和DGGE对沙门氏菌的16S-23S rRNA内部转录间隔区进行了指纹识别。这些技术验证了病原体的克隆性质，但确实显示出区分各种气单胞菌属物种的能力。

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  • 作者

    Quinn, Robert Andrew.;

  • 作者单位

    University of Guelph (Canada).;

  • 授予单位 University of Guelph (Canada).;
  • 学科 Biology Microbiology.
  • 学位 M.Sc.
  • 年度 2008
  • 页码 165 p.
  • 总页数 165
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

  • 入库时间 2022-08-17 11:39:03

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