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https://doi.org/10.15644/asc50/2/2
Comparison of RNA Extraction Methods for Molecular Analysis of Oral Cytology
Mônica Ghislaine Oliveira Alves
; Zavod za bioznanosti i oralnu dijagnostiku Instituta za znanost i tehnologiju (UNESP) Sveučilišta Estadual Paulista, São José dos Campos, São Paulo, Brazil; Medicinski fakultet Sveučilišta Braz Cubas, Mogi das Cruzes, Brazil
Mario Pérez-Sayáns
; Zavod za oralnu medicinu, oralnu kirurgiju i implantologiju Medicinskog i stomatološkog fakulteta, Santiago de Compostela, Španjolska
Maria-Elena Padín-Iruegas
; Područje anatomije i embriologije; Zavod za funkcionalnu biologiju i zdravstvo Sveučilišta Vigo, Pontevedra, Španjolska
Maria Dolores Reboiras-López
; Zavod za oralnu medicinu, oralnu kirurgiju i implantologiju Medicinskog i stomatološkog fakulteta, Santiago de Compostela, Španjolska
José Manuel Suarez-Peñaranda
; Zavod za patologiju i forenzične znanosti Sveučilišne bolnice i Stomatološki fakultet Santiago de Compostela, Santiago de Compostela, La Coruña, Španjolska
Rafael López-López
; Zavod za medicinsku onkologiju Sveučilišne klinike Santiago de Compostela, La Coruña, Španjolska
Celina Faig Lima Carta
; Zavod za bioznanosti i oralnu dijagnostiku Instituta za znanost i tehnologiju (UNESP) Sveučilišta Estadual Paulista, São José dos Campos, São Paulo, Brazil
Jaqueline Scholz Issa
; Program za prestanak pušenja i Područje kardiologije Kardiološkog instituta Sveučilišta São Paulo, Medicinski fakultet i Klinička bolnica, São Paulo, Brazil
Abel García-García
; Zavod za oralnu medicinu, oralnu kirurgiju i implantologiju Medicinskog i stomatološkog fakulteta, Santiago de Compostela, Španjolska
Janete Dias Almeida
; Zavod za bioznanosti i oralnu dijagnostiku Instituta za znanost i tehnologiju (UNESP) Sveučilišta Estadual Paulista, São José dos Campos, São Paulo, Brazil
Sažetak
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.
Ključne riječi
Mouth Mucosa; RNA; Specimen Handling; Molecular Diagnostic Techniques
Hrčak ID:
160214
URI
Datum izdavanja:
20.6.2016.
Posjeta: 2.555 *