Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish
Shewanella algae is an opportunistic pathogen, which is widely distributed in freshwater and marine environments. Diseases caused by S. algae have resulted in substantial economic losses in aquaculture industry. This study aimed to establish a loop mediated isothermal amplification (LAMP) method for...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-10-01
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| Series: | Aquaculture Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352513422002885 |
| _version_ | 1811183752261402624 |
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| author | Zhongjie Zan Zhuoran Han Yuting Hou Yanyu Zhang Jingfeng Sun |
| author_facet | Zhongjie Zan Zhuoran Han Yuting Hou Yanyu Zhang Jingfeng Sun |
| author_sort | Zhongjie Zan |
| collection | DOAJ |
| description | Shewanella algae is an opportunistic pathogen, which is widely distributed in freshwater and marine environments. Diseases caused by S. algae have resulted in substantial economic losses in aquaculture industry. This study aimed to establish a loop mediated isothermal amplification (LAMP) method for rapid and specific detection of S. algae in aquatic animals. The specific LAMP primers were designed according to the 16 S rRNA gene sequences of S. algae strains deposited in GenBank. Based on adjustment of the primer concentration and optimization of reaction conditions, the LAMP method for S. algae detection was established, and specificity and sensitivity tests were carried out. The optimal temperature of this method was 65 °C at constant 1 h. The LAMP amplification reaction was positive for S. algae, but negative for Pseudoalteromonas pisicida, Aeromonas hydrophila, A. veronii, Vibrio parahemolyticus, S. decolorationis, S. xiamenensis, Enterobacter cloacae, Bacillus cereus and Staphylococcus epidermidis. The minimum concentration of template DNA of S. algae for the LAMP method was 8.15 × 10−5 µg/µL. The LAMP method was comparable to the common PCR when they were used for detection of S. algae in koi carp artificially infected with this pathogen. In this study, a rapid, specific and sensitive LAMP method was successfully established for the detection of S. algae, providing great convenience for rapid diagnosis of aquatic animal diseases caused by S. algae infection. |
| first_indexed | 2024-04-11T09:51:08Z |
| format | Article |
| id | doaj.art-d2eddc20bb2e4455abded3643c75b30f |
| institution | Directory Open Access Journal |
| issn | 2352-5134 |
| language | English |
| last_indexed | 2024-04-11T09:51:08Z |
| publishDate | 2022-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Aquaculture Reports |
| spelling | doaj.art-d2eddc20bb2e4455abded3643c75b30f2022-12-22T04:30:47ZengElsevierAquaculture Reports2352-51342022-10-0126101292Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fishZhongjie Zan0Zhuoran Han1Yuting Hou2Yanyu Zhang3Jingfeng Sun4Tianjin key Lab of Aqua-ecology and Aquaculture, Fisheries College, Tianjin Agricultural University, Tianjin 300384, ChinaTianjin key Lab of Aqua-ecology and Aquaculture, Fisheries College, Tianjin Agricultural University, Tianjin 300384, ChinaTianjin key Lab of Aqua-ecology and Aquaculture, Fisheries College, Tianjin Agricultural University, Tianjin 300384, ChinaTianjin key Lab of Aqua-ecology and Aquaculture, Fisheries College, Tianjin Agricultural University, Tianjin 300384, ChinaCorrespondence to: Fisheries College, Tianjin Agricultural University, 22 Jinjing Road, Xiqing District, Tianjin 300384, China.; Tianjin key Lab of Aqua-ecology and Aquaculture, Fisheries College, Tianjin Agricultural University, Tianjin 300384, ChinaShewanella algae is an opportunistic pathogen, which is widely distributed in freshwater and marine environments. Diseases caused by S. algae have resulted in substantial economic losses in aquaculture industry. This study aimed to establish a loop mediated isothermal amplification (LAMP) method for rapid and specific detection of S. algae in aquatic animals. The specific LAMP primers were designed according to the 16 S rRNA gene sequences of S. algae strains deposited in GenBank. Based on adjustment of the primer concentration and optimization of reaction conditions, the LAMP method for S. algae detection was established, and specificity and sensitivity tests were carried out. The optimal temperature of this method was 65 °C at constant 1 h. The LAMP amplification reaction was positive for S. algae, but negative for Pseudoalteromonas pisicida, Aeromonas hydrophila, A. veronii, Vibrio parahemolyticus, S. decolorationis, S. xiamenensis, Enterobacter cloacae, Bacillus cereus and Staphylococcus epidermidis. The minimum concentration of template DNA of S. algae for the LAMP method was 8.15 × 10−5 µg/µL. The LAMP method was comparable to the common PCR when they were used for detection of S. algae in koi carp artificially infected with this pathogen. In this study, a rapid, specific and sensitive LAMP method was successfully established for the detection of S. algae, providing great convenience for rapid diagnosis of aquatic animal diseases caused by S. algae infection.http://www.sciencedirect.com/science/article/pii/S2352513422002885Shewanella algaeLAMP method16 S rRNA geneSpecificitySensitivity |
| spellingShingle | Zhongjie Zan Zhuoran Han Yuting Hou Yanyu Zhang Jingfeng Sun Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish Aquaculture Reports Shewanella algae LAMP method 16 S rRNA gene Specificity Sensitivity |
| title | Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish |
| title_full | Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish |
| title_fullStr | Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish |
| title_full_unstemmed | Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish |
| title_short | Development of a loop-mediated isothermal amplification method for detection of Shewanella algae in fish |
| title_sort | development of a loop mediated isothermal amplification method for detection of shewanella algae in fish |
| topic | Shewanella algae LAMP method 16 S rRNA gene Specificity Sensitivity |
| url | http://www.sciencedirect.com/science/article/pii/S2352513422002885 |
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