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

Karatas S., MIKALSEN J., STEINUM T. M., TAKSDAL T., BORDEVIK M., COLQUHOUN D. J.

JOURNAL OF FISH DISEASES, cilt.31, sa.10, ss.747-753, 2008 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 31 Sayı: 10
  • Basım Tarihi: 2008
  • Doi Numarası: 10.1111/j.1365-2761.2008.00948.x
  • Dergi Adı: JOURNAL OF FISH DISEASES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.747-753
  • Anahtar Kelimeler: detection, embedded tissues, Piscirickettsia salmonis, polymerase chain reaction, RICKETTSIA-LIKE ORGANISM, FARMED ATLANTIC COD, DISEASE, IDENTIFICATION, CONFIRMATION, ASSAYS, SALAR
  • İstanbul Üniversitesi Adresli: Evet

Özet

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.