Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay

Publication: Research - peer-reviewJournal article – Annual report year: 2011

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Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay. / Lievens, B.; Frans, I.; Heusdens, C.; Justé, A.; Jonstrup, Søren Peter; Lieffrig, F.; Willems, K. A.

In: Journal of Fish Diseases, Vol. 34, No. 11, 2011, p. 861-875.

Publication: Research - peer-reviewJournal article – Annual report year: 2011

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Lievens, B.; Frans, I.; Heusdens, C.; Justé, A.; Jonstrup, Søren Peter; Lieffrig, F.; Willems, K. A. / Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay.

In: Journal of Fish Diseases, Vol. 34, No. 11, 2011, p. 861-875.

Publication: Research - peer-reviewJournal article – Annual report year: 2011

Bibtex

@article{66f102235a6147858982fed697513ebb,
title = "Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay",
publisher = "Wiley-Blackwell Publishing Ltd.",
author = "B. Lievens and I. Frans and C. Heusdens and A. Justé and Jonstrup, {Søren Peter} and F. Lieffrig and Willems, {K. A.}",
year = "2011",
doi = "10.1111/j.1365-2761.2011.01304.x",
volume = "34",
number = "11",
pages = "861--875",
journal = "Journal of Fish Diseases",
issn = "0140-7775",

}

RIS

TY - JOUR

T1 - Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay

A1 - Lievens,B.

A1 - Frans,I.

A1 - Heusdens,C.

A1 - Justé,A.

A1 - Jonstrup,Søren Peter

A1 - Lieffrig,F.

A1 - Willems,K. A.

AU - Lievens,B.

AU - Frans,I.

AU - Heusdens,C.

AU - Justé,A.

AU - Jonstrup,Søren Peter

AU - Lieffrig,F.

AU - Willems,K. A.

PB - Wiley-Blackwell Publishing Ltd.

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

KW - Diagnosis

KW - Herpesvirus

KW - Flavobacterium

KW - Multiplex

KW - Koi herpesvirus (KHV)

U2 - 10.1111/j.1365-2761.2011.01304.x

DO - 10.1111/j.1365-2761.2011.01304.x

JO - Journal of Fish Diseases

JF - Journal of Fish Diseases

SN - 0140-7775

IS - 11

VL - 34

SP - 861

EP - 875

ER -