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SPIDIA-DNA: An External Quality Assessment for the pre-analytical phase of blood ...



Malentacchi, F., Pazzagli, M., Simi, L., Orlando, C., Wyrich, R., Hartmann, C. C., Verderio, P., Pizzamiglio, S., Ciniselli, C. M., Tichopad, A., Kubista, M., Gelmini, S. SPIDIA-DNA: An External Quality Assessment for the pre-analytical phase of blood samples used for DNA-based analyses. Clinica Chimica Acta 424: 274–286, 2013. ISSN 0009-8981.

Abstract

Background

The EC-funded project SPIDIA is aimed to develop evidence-based quality guidelines for the pre-analytical phase of blood samples used for DNA molecular testing. To this purpose, a survey and a pan-European External Quality Assessment (EQA) were implemented.

Methods

SPIDIA facility sent to all the participants the same blood sample to be processed without time or temperature limitation. DNA quality parameters performed at SPIDIA facility included: UV spectrophotometric analysis of DNA purity and yield, PCR interferences study by Kineret software and DNA integrity analysis by pulsed field gel electrophoresis.

Results

197 applications have been collected from 30 European countries. A high variability of DNA fragmentation was observed whereas purity, yield and PCR interferences had a narrow distribution within laboratories. A significant difference between the RNase P single copy gene quantity obtained in the DNA samples extracted with the precipitation-based method respect to those obtained with beads and column-based methods was observed.

Conclusions

The results of this study will be the basis for implementing a second pan-European EQA and the results of both EQAs will be pooled and will provide the basis for the implementation of evidence-based guidelines for the pre-analytical phase of DNA analysis of blood samples.