eISSN: 1689-1716
ISSN: 0324-8267
Archiwum Medycyny Sądowej i Kryminologii/Archives of Forensic Medicine and Criminology
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4/2016
vol. 66
 
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abstract:
Original paper

RNA isolation from bloodstains collected on FTA cards – application in clinical and forensic genetics

Katarzyna Skonieczna
1
,
Jan Styczyński
2
,
Anna Krenska
2
,
Mariusz Wysocki
2
,
Aneta Jakubowska
1
,
Tomasz Grzybowski
1

  1. Department of Forensic Medicine, Ludwik Rydygier Collegium Medicum, Nicolaus Copernicus University in Torun, Bydgoszcz
  2. Chair of Pediatrics, Hematology and Oncology, Ludwik Rydygier Collegium Medicum, Nicolaus Copernicus University in Torun, Bydgoszcz
Arch Med Sąd Kryminol 2016; 66 (4): 244–254
DOI: https://doi.org/10.5114/amsik.2016.66706
Online publish date: 2017/06/08
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Aim of the study: In recent years, RNA analysis has been increasingly used in clinical and forensic genetics. Nevertheless, a major limitation of RNA-based applications is very low RNA stability in biological material, due to the RNAse activity. This highlights the need for improving the methods of RNA collection and storage. Technological approaches such as FTA Classic Cards (Whatman) could provide a solution for the problem of RNA degradation. However, different methods of RNA isolation from FTA cards could have diverse effects on RNA quantity and quality. The purpose of this research was to analyze the utility of three different methods of RNA isolation from peripheral blood collected on FTA Classic Cards (Whatman). The study also aimed at assessing RNA stability in bloodstains deposited on FTA cards.

Material and methods: The study was performed on peripheral bloodstains collected from 59 individuals on FTA Classic Cards (Whatman). RNA was isolated with High Pure RNA Isolation Kit (Roche Diagnostics), Universal RNA/miRNA Purification (EURx) and TRIzol Reagent (Life Technologies). RNA was subjected to quantitative analysis followed by reverse transcription and Real – Time PCR reaction.

Results: The study has shown that FTA Classic Cards (Whatman) are useful tools for storing bloodstains at room temperature for RNA analysis. Moreover, the method of RNA extraction employing TRIzol Reagent (Life Technologies) provides the highest efficiency and reproducibility for samples stored for no more than 2 years.

Conclusions: The FTA cards are suitable for collecting and storing bloodstains for RNA analysis in clinical and forensic genetics.
keywords:

bloodstains, RNA, RNA degradation, gene expression, FTA cards
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