Please use this identifier to cite or link to this item:
https://ir.swu.ac.th/jspui/handle/123456789/13698
ชื่อเรื่อง: | Rapid and sensitive detection of Vibrio harveyi by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to vhhP2 gene |
ผู้แต่ง: | Thongkao K. Longyant S. Silprasit K. Sithigorngul P. Chaivisuthangkura P. |
Keywords: | aquaculture industry bacterium bioassay gene gene expression hybridization isotherm polymerase chain reaction sensitivity analysis temperature effect Bacteria (microorganisms) Decapoda (Crustacea) Vibrio Vibrio campbellii Vibrio harveyi |
วันที่เผยแพร่: | 2015 |
บทคัดย่อ: | Vibrio harveyi is a causative agent of the Vibriosis or luminescent bacterial disease in worldwide aquaculture industry. A reliable assay for identification of V. harveyi infection is important to prevent the bacterial spread. In this study, biotinylated loop-mediated isothermal amplification (LAMP) amplicons were produced by a set of four designed primers that recognized specifically the V. harveyi vhhP2 gene, encoding a putative outer membrane protein with unknown function, followed by hybridization with an fluorescein isothiocyanate (FITC)-labelled probe and lateral flow dipstick (LFD) detection. A novel set of PCR primer was also designed specifically to vhhP2 gene and appear to be a species-specific tool for V. harveyi detection. The optimized time and temperature conditions for the LAMP assay were 90 min at 65°C. The LAMP-LFD and PCR methods accurately identified 22 isolates of V. harveyi but did not detect 16 non-harveyi Vibrio isolates, and 34 non-Vibrio bacterial isolates. The sensitivity of LAMP-LFD for V. harveyi detection in pure culture was 1.1 × 102 CFU mL-1 or equivalent to 0.6 CFU per reaction, while that of PCR was 6 CFU per reaction. For spiked shrimp sample, the sensitivity of LAMP was 1.8 × 103 CFU g-1 or equivalent to 5 CFU per reaction, while that of PCR was 50 CFU per reaction. In conclusion, the established LAMP-LFD methods provided a valuable tool for rapid identification of V. harveyi and can be used to distinguish V. harveyi from V. campbellii. © 2013 John Wiley & Sons Ltd. |
URI: | https://ir.swu.ac.th/jspui/handle/123456789/13698 https://www.scopus.com/inward/record.uri?eid=2-s2.0-84926418226&doi=10.1111%2fare.12266&partnerID=40&md5=f4ae4a5f1624a70c589c5f92b1dd2379 |
ISSN: | 1355557X |
Appears in Collections: | Scopus 1983-2021 |
Files in This Item:
There are no files associated with this item.
Items in SWU repository are protected by copyright, with all rights reserved, unless otherwise indicated.