title: Beyond H&E: integration of nucleic acid-based analyses into diagnostic pathology.
abstract: Veterinary pathology of infectious, particularly viral, and neoplastic diseases has advanced significantly with the advent of newer molecular methodologies that can detect nucleic acid of infectious agents within microscopic lesions, differentiate neoplastic from nonneoplastic cells, or determine the suitability of a targeted therapy by detecting specific mutations in certain cancers. Polymerase chain reaction-based amplification of DNA or RNA and in situ hybridization are currently the most commonly used methods for nucleic acid detection. In contrast, the main methodology used for protein detection within microscopic lesions is immunohistochemistry. Other methods that allow for analysis of nucleic acids within a particular cell type or individual cells, such as laser capture microdissection, are also available in some laboratories. This review gives an overview of the factors that influence the accurate analysis of nucleic acids in formalin-fixed tissues, as well as of different approaches to detect such targets.
authors.affiliation: College of Veterinary Medicine, Michigan State University, 4125 Beaumont Road, Lansing, MI 48910, USA. Email: kiupel@dcpah.msu.edu and maes@dcpah.msu.edu.
authors.forename: R K
authors.initials: RK
authors.lastname: Maes
authors.forename: I M
authors.initials: IM
authors.lastname: Langohr
authors.forename: A G
authors.initials: AG
authors.lastname: Wise
authors.forename: R C
authors.initials: RC
authors.lastname: Smedley
authors.forename: T
authors.initials: T
authors.lastname: Thaiwong
authors.forename: M
authors.initials: M
authors.lastname: Kiupel
mesh_terms: D000820:Animal Diseases / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D000818:Animals; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification*; D005557:Formaldehyde / Q000009:adverse effects; D007150:Immunohistochemistry / Q000662:veterinary; D017403:In Situ Hybridization / Q000662:veterinary; D061346:Laser Capture Microdissection / Q000662:veterinary; D009154:Mutation; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D057089:Pathology, Molecular / Q000379:methods*; D010341:Pathology, Veterinary / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011379:Prognosis; D016707:Tissue Fixation / Q000662:veterinary; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary*
chemical_list: D004279:DNA, Viral; D005557:Formaldehyde
keywords: IHC; ISH; PCR; cancer; molecular pathology; nucleic acid analysis; virology