Liquid-based cytology in DNA-based molecular research: viability and potential application

Area of Science:

• Molecular Biology
• Genetics
• Cytology

Background:

• Exfoliated oral cells are a valuable source for non-invasive molecular diagnostics.
• Standardized preservation methods are crucial for reliable DNA analysis from cytologic samples.

Purpose of the Study:

• To assess the efficacy of Universal Collection Medium (UCM) and PreservCyt liquid media for preserving oral exfoliated cells.
• To evaluate the suitability of DNA extracted from these preserved cells for molecular analyses, specifically Tp53 codon 72 polymorphism.

Main Methods:

• Exfoliated oral cells were collected and preserved in UCM and PreservCyt media.
• DNA was extracted using a proteinase K/phenol-chloroform method.
• DNA quantity and quality were assessed via Nanodrop.
• Tp53 codon 72 polymorphism and GADPH genes were analyzed using PCR-RFLP and multiplex PCR, respectively.

Main Results:

• Successful DNA isolation was achieved from all samples.
• Average DNA concentrations were 56.52 ± 34.68 ng/μL (UCM) and 42.13 ± 25.50 ng/μL (PreservCyt).
• Tp53 codon 72 genotyping revealed Pro/Pro (4%), Arg/Arg (60%), and Arg/Pro (36%) genotypes; GADPH gene analysis yielded expected PCR fragments.

Conclusions:

• Both UCM and PreservCyt liquid media are suitable for preserving oral cell samples for subsequent DNA isolation.
• These media facilitate reliable molecular analyses, including gene polymorphism studies.
• Cytologic liquid media represent dependable tools for future research endeavors.