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Title: | Rapid and sensitive detection of Vibrio alginolyticus by loop-mediated isothermal amplification combined with a lateral flow dipstick targeted to the rpoX gene |
Authors: | Plaon S. Longyant S. Sithigorngul P. Chaivisuthangkura P. |
Keywords: | Animalia Bacteria (microorganisms) Litopenaeus vannamei Vibrio Vibrio alginolyticus bacterial protein molecular probe chemistry gene expression regulation genetics isolation and purification metabolism molecular probe nucleic acid amplification physiology procedures Vibrio alginolyticus Bacterial Proteins Gene Expression Regulation, Bacterial Molecular Probes Nucleic Acid Amplification Techniques Vibrio alginolyticus |
Issue Date: | 2015 |
Abstract: | Vibrio alginolyticus is a major bacterial pathogen causing disease in marine animals. The present study aimed to develop a loop-mediated isothermal amplification (LAMP) coupled with a lateral flow dipstick (LFD) for rapid and simple visual detection of V. alginolyticus–specific amplicons. The biotin-labeled LAMP amplicons from the targeted portion of a gene encoding rpoS-like sigma factor (rpoX) were generated at 60°C for 1 h and then hybridized with a fluorescein isothiocyanate–labeled probe for 5 min for visual detection with LFD. In pure cultures, the detection limit of the LAMP–LFD technique for V. alginolyticus was 1.8 × 102 CFU/mL while that of PCR was 1.8 × 103 CFU/mL. In spiked whiteleg shrimp samples Penaeus vannamei, the sensitivity for V. alginolyticus detection was 2 × 103 CFU/g (equivalent to 4 CFU per reaction) while PCR was 10 times less sensitive. The LAMP–LFD method for V. alginolyticus correctly identified 21 isolates of V. alginolyticus but did not recognize 23 non-V. alginolyticus Vibrio isolates and 15 non-Vibrio isolates. In summary, this LAMP–LFD method targeted to the rpoX gene is a convenient assay for specific identification of V. alginolyticus with high sensitivity. © American Fisheries Society 2015. |
URI: | https://ir.swu.ac.th/jspui/handle/123456789/13779 https://www.scopus.com/inward/record.uri?eid=2-s2.0-84940045331&doi=10.1080%2f08997659.2015.1037468&partnerID=40&md5=bf42cede0883a05711c8c7de7b78d58e |
ISSN: | 8997659 |
Appears in Collections: | Scopus 1983-2021 |
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