Journal article
Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay : DNA array-based multiplex assay for fish pathogens
KU Leuven1
Section of Fish Diseases, Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark2
Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark3
National Veterinary Institute, Technical University of Denmark4
Fish Diseases Laboratory5
Fish diseases can be caused by a variety of diverse organisms, including bacteria, fungi, viruses and protozoa, and pose a universal threat to the ornamental fish industry and aquaculture. The lack of rapid, accurate and reliable means by which fish pathogens can be detected and identified has been one of the main limitations in fish pathogen diagnosis and fish disease management and has consequently stimulated the search for alternative diagnostic techniques.
Here, we describe a method based on multiplex and broad‐range PCR amplification combined with DNA array hybridization for the simultaneous detection and identification of all cyprinid herpesviruses (CyHV‐1, CyHV‐2 and CyHV‐3) and some of the most important fish pathogenic Flavobacterium species, including F. branchiophilum, F. columnare and F. psychrophilum.
For virus identification, the DNA polymerase and helicase genes were targeted. For bacterial identification, the ribosomal RNA gene was used. The developed methodology permitted 100% specificity for the identification of the target species. Detection sensitivity was equivalent to 10 viral genomes or less than a picogram of bacterial DNA.
The utility and power of the array for sensitive pathogen detection and identification in complex samples such as infected tissue is demonstrated in this study.
Language: | English |
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Year: | 2011 |
Pages: | 861-875 |
ISSN: | 13652761 and 01407775 |
Types: | Journal article |
DOI: | 10.1111/j.1365-2761.2011.01304.x |
Animals Bacterial Infections Carps DNA Helicases DNA-Directed DNA Polymerase Fish Diseases Fisheries Flavobacteriaceae Infections Herpesviridae Herpesviridae Infections Multiplex Polymerase Chain Reaction Oligonucleotide Array Sequence Analysis RNA, Ribosomal, 16S Reproducibility of Results Sensitivity and Specificity