Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria
文献类型：期刊论文
| 作者 | Zhang, D. F.1,2; Zhang, Q. Q.2; Li, A. H.2 |
| 刊名 | LETTERS IN APPLIED MICROBIOLOGY
 |
| 出版日期 | 2014-11-01 |
| 卷号 | 59期号:5页码:471-478 |
| 关键词 | aquaculture detection fish pathogenic bacteria genus-specific primers multiplex PCR |
| ISSN号 | 0266-8254 |
| 英文摘要 | Species of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus are the most common fish pathogenic bacteria that cause economically devastating losses in aquaculture. A multiplex polymerase chain reaction (mPCR) was developed for the simultaneous detection and differentiation of the four genera of fish pathogenic bacteria. Through the use of genus-specific primers instead of species-specific ones, the current mPCR covered much more target bacterial species compared with previously reported species-specific mPCR methods. The specificity of the four putative genus-specific primers was validated experimentally while used exclusively (uniplex PCR) or combined (mPCR) against bacterial genomic DNA templates of the target bacteria and nontarget bacteria. The PCR amplicons for the following genera were obtained as expected: Aeromonas (875bp), Vibrio (524bp), Edwardsiella (302bp) and Streptococcus (197bp), and the fragments could be separated clearly on the agarose gel electrophoresis. The mPCR did not produce nonspecific amplification products when used to amplify 21 nontarget species of bacteria. The mPCR detection limits for each target bacterial genera were 50 colony-forming units (CFU) in pure culture and 100CFU in fish tissue samples. In conclusion, the mPCR assay was proven to be a powerful alternative to the conventional culture-based method, given its rapid, specific, sensitive and reliable detection of target pathogens. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology ; Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology ; Microbiology |
| 关键词[WOS] | POLYMERASE-CHAIN-REACTION ; STREPTOCOCCUS-AGALACTIAE ; EDWARDSIELLA-ICTALURI ; IN-VITRO ; AEROMONAS ; CHINA ; SEQUENCES ; STRAINS ; TILAPIA ; TARDA |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000343871400003 |
| 公开日期 | 2015-01-20 |
| 源URL | [http://ir.ihb.ac.cn/handle/342005/20329]  |
| 专题 | 水生生物研究所_鱼类生物学及渔业生物技术研究中心_期刊论文 |
| 作者单位 | 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Guangzhou, Guangdong, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, D. F.,Zhang, Q. Q.,Li, A. H.. Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria[J]. LETTERS IN APPLIED MICROBIOLOGY,2014,59(5):471-478. |
| APA | Zhang, D. F.,Zhang, Q. Q.,&Li, A. H..(2014).Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria.LETTERS IN APPLIED MICROBIOLOGY,59(5),471-478. |
| MLA | Zhang, D. F.,et al."Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria".LETTERS IN APPLIED MICROBIOLOGY 59.5(2014):471-478. |
入库方式： OAI收割
来源：水生生物研究所
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