Influence of pre-analytical procedures on genomic DNA integrity in blood samples: the SPIDIA experience

Clin Chim Acta. 2015 Feb 2;440:205-10. doi: 10.1016/j.cca.2014.12.004. Epub 2014 Dec 6.

Abstract

Background: DNA integrity is a critical part of the definition of genomic DNA (gDNA) quality and can influence downstream molecular applications. Pre-analytical variables as sample storage and DNA extraction methods can influence the quality and quantity of isolated DNA and affect molecular test performances. The aim of this paper is to investigate the role of blood sample storage and DNA extraction procedures on gDNA integrity and gDNA fragmentation impact on a molecular test.

Methods: 157 DNA samples deriving from the Pan European 1st SPIDIA DNA External Quality Assessment (EQA), aimed to investigate the influence of blood storage on gDNA quality and quantity, have been analyzed by Pulsed Field Gel Electrophoresis and ImageJ imaging software. 157 DNA samples derived from the Pan European 1st SPIDIA DNA External Quality Assessment (EQA), which aimed to investigate the influence of blood storage on gDNA quality and quantity, have been analyzed by Pulsed Field Gel Electrophoresis and ImageJ imaging software.

Results/conclusions: Our results demonstrate that blood sample storage and DNA extraction procedures influence gDNA integrity and that the same blood sample which underwent a long range multiplex PCR based analytical test can provide different results if the adopted pre-analytical procedures are not standardized.

Keywords: Blood samples; External Quality Assessment; High Molecular Weight gDNA; Pre-analytical phase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Specimen Collection / methods*
  • Chemical Fractionation
  • DNA / blood*
  • DNA / isolation & purification
  • DNA Fragmentation
  • Electrophoresis, Gel, Pulsed-Field
  • Genetic Techniques / standards
  • Humans
  • Molecular Weight
  • Multiplex Polymerase Chain Reaction
  • Quality Control
  • Software

Substances

  • DNA