Original scientific paper
https://doi.org/10.15644/asc50/2/2
Comparison of RNA Extraction Methods for Molecular Analysis of Oral Cytology
Mônica Ghislaine Oliveira Alves
; Department of Biosciences and Oral Diagnosis, Institute of Science and Technology, UNESP - Univ Estadual Paulista, São José dos Campos, São Paulo, Brazil; School of Dentistry, Universidade Braz Cubas, Mogi das Cruzes, Brazil
Mario Pérez-Sayáns
; Oral Medicine, Oral Surgery and Implantology Unit, Faculty of Medicine and Dentistry, Santiago de Compostela, Spain
Maria-Elena Padín-Iruegas
; Human Anatomy and Embriology Area; Functional Biology and Health Sciences Department. University of Vigo, Pontevedra, Spain
Maria Dolores Reboiras-López
; Oral Medicine, Oral Surgery and Implantology Unit, Faculty of Medicine and Dentistry, Santiago de Compostela, Spain
José Manuel Suarez-Peñaranda
; Department of Pathology, Forensic Sciences, University Hospital and School of Medicine of Santiago de Compostela, Santiago de Compostela, La Coruña Spain
Rafael López-López
; Department of Medical Oncology Hospital Clinico Universitario of Santiago de Compostela, La Coruña, Spain
Celina Faig Lima Carta
; Department of Biosciences and Oral Diagnosis, Institute of Science and Technology, UNESP - Univ Estadual Paulista, São José dos Campos, São Paulo, Brazil.
Jaqueline Scholz Issa
; Smoking Cessation Program, Area of Cardiology, Heart Institute, University of São Paulo, School of Medicine, Hospital das Clínicas, São Paulo, Brazil
Abel García-García
; Oral Medicine, Oral Surgery and Implantology Unit, Faculty of Medicine and Dentistry, Santiago de Compostela, Spain
Janete Dias Almeida
; Department of Biosciences and Oral Diagnosis, Institute of Science and Technology, UNESP - Univ Estadual Paulista, São José dos Campos, São Paulo, Brazil.
Abstract
Objective of work: The aim of this study was to compare three methods of RNA extraction for molecular analysis of oral cytology to establish the best technique, considering its concentration and purity
for molecular tests of oral lesions such as real-time reverse transcriptase reaction. Material and methods: The sample included exfoliative cytology from the oral cavity mucosa of patients with no
visible clinical changes, using Orcellex Rovers Brush®. The extraction of total RNA was performed using the following three techniques: 30 samples were extracted by Trizol® technique, 30 by the DirectzolTM
RNA Miniprep system and 30 by the RNeasy mini Kit. The absorbance was measured by spectrophotometer to estimate the purity. The estimated RNA concentration was obtained by multiplying the value of A260 (ng/mL) by 40. Statistical analysis of the obtained data was performed using GraphPad Prism 5.03 software with Student t, analysis of variance and Bonferroni tests, considering p ≤0.05. Results: Trizol® group revealed higher average concentration, followed by Direct-zolTM and Rneasy group. It was observed that the RNA Direct-zolTM group had the highest purity, followed by RNeasy
and Trizol® groups, allowing for the two ratios. Conclusion: Considering all aspects, concentration, purity and time spent in the procedures, the Direct-zolTM group showed the best results.
Keywords
Mouth Mucosa; RNA; Specimen Handling; Molecular Diagnostic Techniques
Hrčak ID:
160214
URI
Publication date:
20.6.2016.
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