Investigation of DNA and RNA viruses from paraffin-embedded tissue blocks using molecular methods
Öz
Polymerase chain reaction (PCR) is the most used molecular method in routine diagnosis for viral diseases. It is chosen for its high sensitivity and rapid results. One of the most important factors for reliability of results in PCR is the choice of sample material. Cold chain is mandatory for transporting the material and organ samples in particular may start decomposing if conditions are compromised and transport time is prolonged. Field staff may decide not to send organ samples in a case like this. Alternatively, formalin fixed paraffin embedded tissues do not need any specific conditions in transport which poses an advantage. The aim of this study is to create awareness in value of paraffin blocks in molecular diagnostics and investigate their practical importance in PCR applications. Two different frequently studied viruses were chosen, bovine papillomavirus (BPV) is chosen for representing DNA viruses and canine distemper virus (CDV) is chosen for representing RNA viruses. Five paraffin embedded tissue blocks for each virus, which are previously known to be positive, were used as material. Two different sets of primer pairs were chosen for PCR application, MY09/MY11 targeting L1 ORF region for BPV and a primer pair targeting nucleocapsid region for CDV. Reverse transcription was performed for samples of CDV before the PCR application. Four of the BPV samples and two of the CDV samples were deemed positive following PCR and RT-PCR. According to the results, paraffin embedded tissue blocks can be regarded valuable, albeit more efficiently for DNA viruses and less efficiently in RNA viruses. Additionally, our findings indicate there might be false negative results and this should be taken into consideration when planning the studies.
Anahtar Kelimeler
Virus, Polymerase Chain Reaction, Paraffin Embedded Tissue Block
Kaynakça
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