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A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
Published on: March 30, 2020
Detection of alpha human papillomaviruses in archival formalin-fixed, paraffin-embedded (FFPE) tissue specimens
Boštjan J Kocjan1, Lea Hošnjak1, Mario Poljak2
1Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
Abstract:
Formalin-fixed, paraffin-embedded (FFPE) tissue specimens stored in pathology departments worldwide are an invaluable source for diagnostic purposes when fresh clinical material is unavailable as well as for retrospective molecular and epidemiological studies, especially when dealing with rare clinical conditions for which prospective collection is not feasible. Accurate detection of HPV infection in these specimens is particularly challenging because nucleic acids are often degraded and therefore, not suitable for amplification of larger fragments of the viral genome or viral gene transcripts. This review provides a brief summary of molecular methods for detecting alpha-HPV DNA/RNA in FFPE tissue specimens. We specifically address the key procedural and environmental factors that have the greatest impact on the quality of nucleic acids extracted from FFPE tissue specimens, and describe some solutions that can be used to increase their integrity and/or amplifiability. Moreover, commonly used methods for HPV DNA/RNA detection in FFPE tissue specimens are presented and discussed, focusing on studies using polymerase chain reaction as an HPV detection method and published after 1999. Finally, we briefly summarize our 22 years of experience with HPV detection in FFPE tissue specimens.
Insights
Detecting human papillomavirus (HPV) DNA/RNA in formalin-fixed, paraffin-embedded (FFPE) tissues is challenging due to degraded nucleic acids. This review summarizes molecular methods and factors affecting HPV detection in FFPE specimens.
Area of Science:
- Pathology
- Molecular Biology
- Virology
Background:
- Formalin-fixed, paraffin-embedded (FFPE) tissues are crucial for diagnostics and retrospective studies, especially for rare diseases.
- Detecting human papillomavirus (HPV) in FFPE specimens is difficult due to degraded nucleic acids, hindering amplification.
- This review focuses on molecular detection of alpha-HPV DNA/RNA in FFPE tissues.
Purpose of the Study:
- To review molecular methods for HPV DNA/RNA detection in FFPE specimens.
- To identify factors impacting nucleic acid quality and amplifiability in FFPE tissues.
- To discuss polymerase chain reaction (PCR)-based HPV detection methods published since 1999.
Main Methods:
- Review of molecular techniques for HPV DNA/RNA detection in FFPE tissues.
- Analysis of procedural and environmental factors affecting nucleic acid integrity.
- Focus on PCR-based methods for HPV detection in FFPE specimens.
Main Results:
- Nucleic acid degradation in FFPE tissues presents significant challenges for HPV detection.
- Specific procedural and environmental factors critically influence nucleic acid quality.
- Various molecular methods, particularly PCR, are employed for HPV detection in FFPE samples.
Conclusions:
- Effective HPV detection in FFPE tissues requires careful consideration of nucleic acid quality and extraction methods.
- Optimizing protocols can enhance the integrity and amplifiability of nucleic acids from FFPE specimens.
- PCR-based methods remain a primary approach for HPV DNA/RNA detection in FFPE tissues, with ongoing advancements.
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