Real time PCR detection of Piscirickettsia salmonis from formalin-fixed paraffin-embedded tissues.
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.
Karatas
S
S
National Veterinary Institute, Oslo, Norway.
Mikalsen
J
J
Steinum
T M
TM
Taksdal
T
T
Bordevik
M
M
Colquhoun
D J
DJ
D000818:Animals; D005393:Fish Diseases / Q000175:diagnosis*; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D016612:Paraffin Embedding; D053522:Piscirickettsia / Q000302:isolation & purification*; D044225:Piscirickettsiaceae Infections / Q000175:diagnosis / Q000662:veterinary*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D016707:Tissue Fixation
D005404:Fixatives; D005557:Formaldehyde