Real time PCR detection of Piscirickettsia salmonis from formalin-fixed paraffin-embedded tissues.

Abstract

Piscirickettsia salmonis is the causative agent of piscirickettsiosis, a transmissible disease of salmonid fish. Diagnosis of piscirickettsiosis has traditionally been based upon identification of typical pathological changes by histological investigation, with confirmation by immunohistochemistry on formalin-fixed, paraffin-embedded tissues. However, implementation of more rapid confirmatory techniques, preferably with higher levels of sensitivity and possibilities for quantification, is desirable. A real-time polymerase chain reaction (PCR) assay was designed for specific detection of P. salmonis and tested on samples extracted from formalin-fixed paraffin-embedded material. Construction of a PCR-target mimic allowed determination of detection limits, linearity of the real-time PCR and quantitative detection of P. salmonis. The present study demonstrates the capability of the described real time PCR assay for detection of P. salmonis from paraffin-embedded material with a high degree of sensitivity and specificity. Implementation of this assay constitutes an important development for a rapid and secure diagnosis of piscirickettsiosis.

• Authors: S Karatas, J Mikalsen, T M Steinum, T Taksdal, M Bordevik, D J Colquhoun
• Journal: Journal of fish diseases (J Fish Dis) Vol. 31 Issue 10 Pg. 747-53 (Oct 2008) ISSN: 1365-2761 [Electronic] England
• PMID: 18681901 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Fixatives
• Formaldehyde

Topics

• Animals
• Fish Diseases (diagnosis)
• Fixatives (chemistry)
• Formaldehyde (chemistry)
• Paraffin Embedding
• Piscirickettsia (isolation & purification)
• Piscirickettsiaceae Infections (diagnosis, veterinary)
• Polymerase Chain Reaction
• Reproducibility of Results
• Sensitivity and Specificity
• Time Factors
• Tissue Fixation