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Development of sensitive, high-throughput one-tube RT-PCR-enzyme hybridisation assay to detect selected bacterial fish pathogens

Citation

Wilson, TK and Carson, J, Development of sensitive, high-throughput one-tube RT-PCR-enzyme hybridisation assay to detect selected bacterial fish pathogens, Diseases of Aquatic Organisms, 54, (2) pp. 127-134. ISSN 0177-5103 (2003) [Refereed Article]

DOI: doi:10.3354/dao054127

Abstract

Bacterial monitoring and surveillance is critical for the early detection of pathogens to avoid the spread of disease. To facilitate this, an efficient, high-performance and high-throughput method to detect the presence of femotgram amounts of ribosomal RNA from 4 bacterial fish pathogens: Aeromonas salmonicida; Tenacibaculum maritimum (formerly Flexibacter maritimus); Lactococcus garvieae; and Yersinia ruckeri was developed. The system uses NucleoLink™ strips for liquid- and solid-phase PCR in 1 tube, to perform RT-PCR-enzyme hybridisation assays (RT-PCR-EHA) detecting 4 fg or less of rRNA from pure cultures and between 1 and 9 CFU per 200 μl sample volume from selective-enrichment culture media. The liquid-phase amplicons were visualised by gel electrophoresis and the solid-phase amplicons detected using internal probes and visualised using colorimetric detection and p-nitrophenylphosphate.

Item Details

Item Type:Refereed Article
Research Division:Agricultural and Veterinary Sciences
Research Group:Fisheries Sciences
Research Field:Fish Pests and Diseases
Objective Division:Environment
Objective Group:Control of Pests, Diseases and Exotic Species
Objective Field:Control of Animal Pests, Diseases and Exotic Species in Coastal and Estuarine Environments
Author:Wilson, TK (Ms Teresa Wilson)
Author:Carson, J (Dr Jeremy Carson)
ID Code:29058
Year Published:2003
Web of Science® Times Cited:16
Deposited By:TAFI - Fish Health Unit
Deposited On:2003-08-01
Last Modified:2004-06-21
Downloads:0

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