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Evaluation of Cytologic Sample Preparations for Compatibility With Nucleic Acid Analysis
Laure Sorber1, Bart Claes2,3, Karen Zwaenepoel1,4
1Center for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), Wilrijk, Belgium.
American Journal of Clinical Pathology
|September 20, 2021
Summary
Alcohol-based collection media best preserve nucleic acids (DNA and RNA) in cytologic samples. Specimen type and processing impact quality, but staining is not detrimental, and formalin-fixed, paraffin-embedded blocks are usable despite reduced quality.
Area of Science:
- Cytopathology
- Molecular Diagnostics
- Biomarker Analysis
Background:
- Cytologic sample workflow significantly impacts nucleic acid quality.
- Optimizing sample handling is crucial for accurate molecular testing.
Purpose of the Study:
- To evaluate the influence of key cytologic workflow elements on DNA and RNA content.
- To identify optimal pre-analytical variables for molecular analysis of cytologic specimens.
Main Methods:
- Utilized A549 cell line, organoids, and pleural effusions.
- Investigated collection media, processing delays, specimen types, staining, and FFPE processing.
- Assessed nucleic acid quality and quantity using fragment analysis, Qubit, and qPCR on the Idylla platform.
Main Results:
- Alcohol-based media (CytoRich Red, EtOH95%) showed high DNA/RNA preservation.
- Phosphate-buffered saline and formalin favored RNA quality.
- Cytospin and smear specimens exhibited nucleic acid loss.
- Cytologic staining did not impact quality; FFPE sections showed decreased quality but remained compatible.
Conclusions:
- Key cytologic workflow steps influence DNA/RNA quality and quantity.
- Selection of workflow elements depends on downstream testing requirements.
