Fluorescent In Situ hybridization: a new tool for the direct identification and detection of F. psychrophilum

Strepparava, N.; Wahli, Thomas; Segner, Helmut; Polli, B.; Petrini, O. (2012). Fluorescent In Situ hybridization: a new tool for the direct identification and detection of F. psychrophilum. PLoS ONE, 7(11), e49280. Public Library of Science 10.1371/journal.pone.0049280

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F. psychrophilum is the causative agent of Bacterial Cold Water Disease (BCW) and Rainbow Trout Fry Syndrome (RTFS). To date, diagnosis relies mainly on direct microscopy or cultural methods. Direct microscopy is fast but not very reliable, whereas cultural methods are reliable but time-consuming and labor-intensive. So far fluorescent in situ hybridization (FISH) has not been used in the diagnosis of flavobacteriosis but it has the potential to rapidly and specifically detect F. psychrophilum in infected tissues. Outbreaks in fish farms, caused by pathogenic strains of Flavobacterium species, are increasingly frequent and there is a need for reliable and cost-effective techniques to rapidly diagnose flavobacterioses. This study is aimed at developing a FISH that could be used for the diagnosis of F. psychrophilum infections in fish. We constructed a generic probe for the genus Flavobacterium ("Pan-Flavo") and two specific probes targeting F. psychrophilum based on 16S rRNA gene sequences. We tested their specificity and sensitivity on pure cultures of different Flavobacterium and other aquatic bacterial species. After assessing their sensitivity and specificity, we established their limit of detection and tested the probes on infected fresh tissues (spleen and skin) and on paraffin-embedded tissues. The results showed high sensitivity and specificity of the probes (100% and 91% for the Pan-Flavo probe and 100% and 97% for the F. psychrophilum probe, respectively). FISH was able to detect F. psychrophilum in infected fish tissues, thus the findings from this study indicate this technique is suitable as a fast and reliable method for the detection of Flavobacterium spp. and F. psychrophilum.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Center for Fish and Wildlife Health (FIWI)

UniBE Contributor:

Wahli, Thomas and Segner, Helmut

Subjects:

600 Technology > 630 Agriculture

ISSN:

1932-6203

Publisher:

Public Library of Science

Language:

English

Submitter:

Andrea Stettler

Date Deposited:

27 Jan 2014 11:15

Last Modified:

13 Dec 2014 10:26

Publisher DOI:

10.1371/journal.pone.0049280

PubMed ID:

23152887

Uncontrolled Keywords:

Animals, DNA Probes/metabolism, DNA, ibosomal/genetics, Fish Diseases/diagnosis/microbiology, Fishes/microbiology, Flavobacteriaceae Infections/diagnosis/microbiology, veterinary, Flavobacterium/*genetics/*isolation & purification, In Situ Hybridization, Fluorescence/*methods, Limit of Detection, Paraffin Embedding, ROC Curve, Sequence Analysis, DNA

BORIS DOI:

10.7892/boris.40452

URI:

https://boris.unibe.ch/id/eprint/40452

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