Detection and quantification of Flavobacterium psychrophilum in water and fish tissue samples by quantitative real time PCR.

Strepparava, Nicole; Wahli, Thomas; Segner, Helmut; Petrini, Orlando (2014). Detection and quantification of Flavobacterium psychrophilum in water and fish tissue samples by quantitative real time PCR. BMC microbiology, 14, p. 105. BioMed Central 10.1186/1471-2180-14-105

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BACKGROUND Flavobacterium psychrophilum is the agent of Bacterial Cold Water Disease and Rainbow Trout Fry Syndrome, two diseases leading to high mortality. Pathogen detection is mainly carried out using cultures and more rapid and sensitive methods are needed. RESULTS We describe a qPCR technique based on the single copy gene β' DNA-dependent RNA polymerase (rpoC). Its detection limit was 20 gene copies and the quantification limit 103 gene copies per reaction. Tests on spiked spleens with known concentrations of F. psychrophilum (106 to 101 cells per reaction) showed no cross-reactions between the spleen tissue and the primers and probe. Screening of water samples and spleens from symptomless and infected fishes indicated that the pathogen was already present before the outbreaks, but F. psychrophilum was only quantifiable in spleens from diseased fishes. CONCLUSIONS This qPCR can be used as a highly sensitive and specific method to detect F. psychrophilum in different sample types without the need for culturing. qPCR allows a reliable detection and quantification of F. psychrophilum in samples with low pathogen densities. Quantitative data on F. psychrophilum abundance could be useful to investigate risk factors linked to infections and also as early warning system prior to potential devastating outbreak.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Veterinary Public Health / Herd Health Management
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Center for Fish and Wildlife Health (FIWI)

UniBE Contributor:

Strepparava, Nicole; Wahli, Thomas and Segner, Helmut

Subjects:

600 Technology > 630 Agriculture

ISSN:

1471-2180

Publisher:

BioMed Central

Language:

English

Submitter:

Lucia Gugger-Raaflaub

Date Deposited:

23 Apr 2015 11:44

Last Modified:

23 Apr 2015 11:44

Publisher DOI:

10.1186/1471-2180-14-105

PubMed ID:

24767577

BORIS DOI:

10.7892/boris.65482

URI:

https://boris.unibe.ch/id/eprint/65482

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