Investigation of DNA and RNA viruses from paraffin-embedded tissue blocks using molecular methods
Journal Title: Antakya Veteriner Bilimleri Dergisi (The Journal of Antakya Veterinary Sciences) - Year 2022, Vol 1, Issue 1
Abstract
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 material. Cold chain is mandatory for transporting the material. Material may start decomposing if conditions are compromised and transport time is prolonged, this is especially important if organs are used as material. 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 viruses were chosen in the design of the study. 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. L1 ORF region of BPV and nucleocapsid region for CDV was targeted. 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 valuable, albeit more efficient for DNA viruses and less efficient in RNA viruses. Additionally, our findings indicated there might be false negative results which should be considered when planning the studies.
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