Biochemistry & Analytical Biochemistry

A novel method for room temperature distribution and conservation of RNA and DNA reference materials for guaranteeing performance of molecular diagnostics in onco-hematology: A GBMHM study

International Conference on Clinical Chemistry & Laboratory Medicine

October 17-18, 2016 Chicago, USA

Jean-Michel Cayuela, Carole Maute, Anne-Lise Fabre, Olivier Nibourel, Stephanie Dulucq, Eric Delabesse, Patrick Villarese, Sandrine Hayette, Marie- Joelle Mozziconacci and Elizabeth Macintyre

University Hospital Saint-Louis, France
University Paris Diderot, France
Imagene, France
Centre Hospitalier Regional Universitaire de Lille, France
University Hospital Bordeaux, France
University Hospital Toulouse, France

Scientific Tracks Abstracts: Biochem Anal Biochem

Abstract:

Objectives: Performance of methods used for molecular diagnostics must be closely controlled by regular analysis of internal quality controls. However, conditioning, shipping and long lasting storage of nucleic acid controls remains problematic. Therefore, we evaluated the minicapsules-based innovative process developed by Imagene (Evry, France) for implementing DNA and RNA controls designed for clonality assessment of lymphoproliferations and BCR-ABL1 mRNA quantification, respectively. Design & Methods: DNA samples were extracted from 12 cell lines selected for giving specific amplifications with most BIOMED-2 PCR tubes. RNA samples were extracted from 8 cell line mixtures expressing various BCR-ABL1 transcript levels. DNA and RNA were encapsulated by Imagene and shipped at room temperature to participating laboratories. Biologists were asked to report quality data of recovered nucleic acids as well as PCR results. Results: encapsulated nucleic acids samples were easily and efficiently recovered from minicapsules. The expected rearrangements at immunoglobulin, T-cell receptor and BCL2 loci were detected in DNA samples by all laboratories. Quality of RNA was consistent between laboratories and met the criteria requested for quantification of BCR-ABL1 transcripts. Expression levels measured by the 5 laboratories were within ±2 fold interval from the corresponding pre-encapsulation reference value. Moreover aging studies of encapsulated RNA simulating up to 100 years storage at room temperature show no bias in quantitative outcome. Conclusions: Therefore, Imagene minicapsules are suitable for storage and distribution at room temperature of genetic material designed for proficiency control of molecular diagnostic methods based on end point or real-time quantitative PCR. We are currently working on novel applications of minicapsules to develop for instance quality controls for DNA extraction in the context of external quality assessment program (EEQ).

Biography :

Jean-Michel Cayuela is an Assistant Professor at University Hospital Saint-Louis in Paris. He has obtained his Medical Biologist Diploma in 1991 and has completed his PhD in 1997. He has been involved in molecular diagnostic in the field of malignant hemopathies for more than 20 years. In this context, he initiated the French external quality control program ten years ago. At the European level he is a Member of the European Leukemia Net (ELN) and of the European Scientific Foundation of Laboratory Hemato Oncology (ESLHO).

Email: jean-michel.cayuela@aphp.fr