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DOI: 10.1371/journal.pone.0170992
论文题名:
Approach to determine the diversity of Legionella species by nested PCR-DGGE in aquatic environments
作者: Wen-Chien Huang; Hsin-Chi Tsai; Chi-Wei Tao; Jung-Sheng Chen; Yi-Jia Shih; Po-Min Kao; Tung-Yi Huang; Bing-Mu Hsu
刊名: PLOS ONE
ISSN: 1932-6203
出版年: 2017
发表日期: 2017-2-6
卷: 12, 期:2
语种: 英语
英文关键词: Legionella ; Legionella pneumophila ; Denaturing gradient gel electrophoresis ; Polymerase chain reaction ; Surface water ; Rivers ; Ribosomal RNA ; DNA sequence analysis
英文摘要: In this study, we describe a nested PCR-DGGE strategy to detect Legionella communities from river water samples. The nearly full-length 16S rRNA gene was amplified using bacterial primer in the first step. After, the amplicons were employed as DNA templates in the second PCR using Legionella specific primer. The third round of gene amplification was conducted to gain PCR fragments apposite for DGGE analysis. Then the total numbers of amplified genes were observed in DGGE bands of products gained with primers specific for the diversity of Legionella species. The DGGE patterns are thus potential for a high-throughput preliminary determination of aquatic environmental Legionella species before sequencing. Comparative DNA sequence analysis of excised DGGE unique band patterns showed the identity of the Legionella community members, including a reference profile with two pathogenic species of Legionella strains. In addition, only members of Legionella pneumophila and uncultured Legionella sp. were detected. Development of three step nested PCR-DGGE tactic is seen as a useful method for studying the diversity of Legionella community. The method is rapid and provided sequence information for phylogenetic analysis.
URL: http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0170992&type=printable
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资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/25788
Appears in Collections:过去全球变化的重建
影响、适应和脆弱性
科学计划与规划
气候变化与战略
全球变化的国际研究计划
气候减缓与适应
气候变化事实与影响

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作者单位: Department of Medicine, Mackay Medicine College, Taipei, Taiwan, ROC;Department of Thoracic Surgery, Mackay Memorial Hospital, Taipei, Taiwan, ROC;Mackay Junior College of Medicine, Nursing, and Management, Taipei, Taiwan, ROC;School of Medicine Tzu-Chi University, Hualien, Taiwan, ROC;Department of Psychiatry, Tzu-Chi General Hospital, Hualien, Taiwan, ROC;Section of Respiratory Therapy, Cheng Hsin General Hospital, Taipei, Taiwan, ROC;Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, ROC;Department of Earth and Environmental Sciences, National Chung Cheng University, Chiayi, Taiwan, ROC;Department of Earth and Environmental Sciences, National Chung Cheng University, Chiayi, Taiwan, ROC;Department of Earth and Environmental Sciences, National Chung Cheng University, Chiayi, Taiwan, ROC;Department of Earth and Environmental Sciences, National Chung Cheng University, Chiayi, Taiwan, ROC

Recommended Citation:
Wen-Chien Huang,Hsin-Chi Tsai,Chi-Wei Tao,et al. Approach to determine the diversity of Legionella species by nested PCR-DGGE in aquatic environments[J]. PLOS ONE,2017-01-01,12(2)
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