期刊
JOURNAL OF FISH DISEASES
卷 31, 期 10, 页码 747-753出版社
WILEY
DOI: 10.1111/j.1365-2761.2008.00948.x
关键词
detection; embedded tissues; Piscirickettsia salmonis; polymerase chain reaction
资金
- NATO Science Fellowship Programme by The Scientific and Technical Research Council of Turkey (TUBITAK)
- Pharmaq AS, Oslo, Norway
- National Veterinary Institute, Oslo, Norway
- Research Fund of Istanbul University, Turkey [UDP-863/07092006]
- The Research Council of Norway [158882]
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.
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