Real time PCR detection of Piscirickettsia salmonis from formalin-fixed paraffin-embedded tissues
JOURNAL OF FISH DISEASES, vol.31, no.10, pp.747-753, 2008 (SCI-Expanded, Scopus)
- Publication Type: Article / Article
- Volume: 31 Issue: 10
- Publication Date: 2008
- Doi Number: 10.1111/j.1365-2761.2008.00948.x
- Journal Name: JOURNAL OF FISH DISEASES
- Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Page Numbers: pp.747-753
- Keywords: detection, embedded tissues, Piscirickettsia salmonis, polymerase chain reaction, RICKETTSIA-LIKE ORGANISM, FARMED ATLANTIC COD, DISEASE, IDENTIFICATION, CONFIRMATION, ASSAYS, SALAR
- Istanbul University Affiliated: Yes
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.