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

Abstract

OBJECTIVE To evaluate the effectiveness of UCM and PreservCyt liquid media in molecular analysis using exfoliated oral cells and analyze Tp53 codon 72 extracted from these cells. STUDY DESIGN Exfoliated oral cells were placed in UCM and PreservCyt preservation media. DNA was isolated by proteinase K/phenol-chloroform protocol. Quantity and quality was measured by Nanodrop 1000. To evaluate the applicability of the isolated DNA to molecular techniques, DNA from oral cells was used to analyze Tp53 codon 72 polymorphism and GADPH genes. RESULTS DNA was successfully isolated from all samples, with an average concentration of 56.52 +/- 34.68 ng/microL for UCM and 42.13 +/- 25.50 ng/microL for PreservCyt. Average 260/280 ratios were 1.83 +/- 0.06 for UCM and 1.74 +/- 0.11 for PreserveCyt. Analysis of Tp53 codon 72 by polymerase chain reaction-restriction fragment length polymorphism was performed for all samples, with 4% of Pro/Pro, 60% of Arg/Arg and 36% of Arg/Pro genotype. In the analysis of GADPH multiplex polymerase chain reaction, fragments varying from 100 to 400 bp were obtained from all samples. CONCLUSION Cytologic liquid media are suitable for preserving cell samples for posterior DNA isolation and molecular analysis and could become reliable research tools.

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