Diagnostic efficacy of molecular assays for the viral haemorrhagic septicaemia virus isolates from the Czech Republic

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The diagnostic properties of the one-step real-time reverse-transcription polymerase chain reaction assay for viral haemorrhagic septicaemia virus detection were compared to methods currently in use in the Czech Republic, namely, virus isolation using the cell culture and conventional reverse-transcription polymerase chain reaction followed by the nested polymerase chain reaction. The assays were tested on a panel of 25 archived viral haemorrhagic septicaemia isolates and 8 archived infectious haematopoietic necrosis isolates obtained from monitoring and/or outbreaks of the diseases among farmed salmonids in the Czech Republic. The ability to detect the presence of the virus in the tissues of fish was tested on additional 32 field samples collected from the rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta) and brook trout (Salvelinus fontinalis). The real-time assay showed the highest analytic sensitivity by detecting the presence of viral nucleic acid in samples with 10*-7 dilution, whereas the sensitivity of the conventional polymerase chain reaction peaked at 10*-5. Diagnostic specificity of both molecular assays was confirmed by absence of cross-reactivity with the infectious haematopoietic necrosis virus isolates. This, along with consistent results in the detection of the virus in the fish tissues, confirms that the one-step real-time reverse-transcription polymerase chain reaction is currently an optimal stand-alone diagnostic method for the detection of the viral haemorrhagic septicaemia virus. … full description

Bibliographic Details

Main Author

Ľubomír Pojezdal

Other Authors

Dagmar Pokorová , Stanislava Reschová, 1964- , Miroslava Palíková, 1966- , Monika Vícenová, Tomáš Veselý , Stanislav Navrátil

Document Type

Articles

Physical Description

Ilustrace

Published in

Acta veterinaria. -- ISSN 0001-7213. -- Roč. 86, č. 3 (2017), s. 207-212

Subjects

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FISHES

lososovití

FISH DISEASES

RHABDOVIRIDAE

FISH RHABDOVIRUSES

HAEMORRHAGIC SEPTICAEMIA VIRUS

Trout hemorrhagic septicemia

LABORATORY DIAGNOSIS

CELL CULTURE

PCR

HAEMATOPOIETIC NECROSIS VIRUS

TROUT

ONCORHYNCHUS MYKISS

SALMO TRUTTA

CHARS

ryby

choroby ryb

infekční choroby

virové nemoci

Rhabdoviridae

rybí rhabdoviry

virus hemorhagické septikémie

hemoragická septikémie pstruha

diagnostické metody

laboratorní diagnóza

buněčná kultura

virus infekční hematopoetické nekrózy

pstruzi

Oncorhynchus mykiss

pstruh obecný

siveni

siven americký

Česko

RT-PCR

more… less…

Item Description

3 tabulky

Bibliography

Literatura na s. 212