Beyond H&E: integration of nucleic acid-based analyses into diagnostic pathology

Area of Science:

• Veterinary pathology
• Molecular diagnostics
• Oncology

Background:

• Advancements in molecular methodologies have significantly improved the diagnosis of infectious and neoplastic diseases in veterinary pathology.
• These techniques enable the detection of nucleic acids from infectious agents and the differentiation of neoplastic from non-neoplastic cells.
• Molecular analysis also aids in determining the suitability of targeted therapies by identifying specific cancer mutations.

Purpose of the Study:

• To provide an overview of molecular methodologies used in veterinary pathology.
• To discuss factors affecting accurate nucleic acid analysis in formalin-fixed tissues.
• To review different approaches for detecting nucleic acid targets in veterinary samples.

Main Methods:

• Polymerase chain reaction (PCR)-based amplification of DNA or RNA for nucleic acid detection.
• In situ hybridization for localized nucleic acid detection within tissue sections.
• Immunohistochemistry for protein detection in microscopic lesions.
• Laser capture microdissection for isolating specific cell types for analysis.

Main Results:

• Molecular methods offer precise detection of infectious agents and cellular abnormalities.
• These techniques are crucial for characterizing neoplastic diseases and guiding therapeutic strategies.
• Accurate analysis of nucleic acids in formalin-fixed tissues is influenced by various pre-analytical and analytical factors.

Conclusions:

• Molecular pathology has revolutionized veterinary diagnostics for infectious and neoplastic diseases.
• The accurate application of techniques like PCR, in situ hybridization, and immunohistochemistry is vital for reliable disease characterization.
• Further understanding of factors influencing nucleic acid analysis will enhance diagnostic accuracy and therapeutic decisions in veterinary medicine.