title: 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.
abstract: OBJECTIVE
  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 remain problematic. Therefore, we evaluated the minicapsule-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.
  
  
  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.
authors: Laboratory of Hematology, University Hospital Saint-Louis, AP-HP, Paris, France; EA3518, University Paris Diderot, Paris, France. Electronic address: jean-michel.cayuela@sls.aphp.fr.
authors: Jean-Michel
authors: JM
authors: Cayuela
authors: Laboratory of Hematology, University Hospital Saint-Louis, AP-HP, Paris, France.
authors: Carole
authors: C
authors: Mauté
authors: Imagene, Evry, France.
authors: Anne-Lise
authors: AL
authors: Fabre
authors: Laboratory of Hematology, CHRU, Lille, France.
authors: Olivier
authors: O
authors: Nibourel
authors: Laboratory of Hematology, University Hospital, Bordeaux, France.
authors: Stéphanie
authors: S
authors: Dulucq
authors: Laboratory of Hematology, University Hospital, Toulouse, France.
authors: Eric
authors: E
authors: Delabesse
authors: Laboratory of Hematology, University Hospital Necker-Enfants-Malades, AP-HP, Paris, France.
authors: Patrick
authors: P
authors: Villarèse
authors: Laboratory of Hematology, University Hospital, Lyon, France.
authors: Sandrine
authors: S
authors: Hayette
authors: Laboratoire de Biopathologie, Institut Paoli Calmettes, Marseille, France.
authors: Marie-Joelle
authors: MJ
authors: Mozziconacci
authors: Laboratory of Hematology, University Hospital Necker-Enfants-Malades, AP-HP, Paris, France.
authors: Elizabeth
authors: E
authors: Macintyre
mesh_terms: D002460:Cell Line; D045744:Cell Line, Tumor; D004247:DNA / Q000032:analysis* / Q000378:metabolism / Q000592:standards; D005240:Feasibility Studies; D005602:France; D016044:Fusion Proteins, bcr-abl / Q000097:blood / Q000235:genetics / Q000378:metabolism; D005820:Genetic Testing / Q000592:standards; D006405:Hematology / Q000379:methods*; D006801:Humans; D059021:Laboratory Proficiency Testing; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood / Q000175:diagnosis / Q000235:genetics; D008232:Lymphoproliferative Disorders / Q000097:blood / Q000175:diagnosis / Q000235:genetics; D008495:Medical Oncology / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000592:standards; D010865:Pilot Projects; D010949:Plasma / Q000737:chemistry; D011786:Quality Control; D012313:RNA / Q000032:analysis* / Q000378:metabolism / Q000592:standards; D020871:RNA Stability; D012333:RNA, Messenger / Q000097:blood / Q000378:metabolism; D012015:Reference Standards; D013696:Temperature
chemical_list: C581634:BCR-ABL1 fusion protein, human; D012333:RNA, Messenger; D012313:RNA; D004247:DNA; D016044:Fusion Proteins, bcr-abl
keywords: Chronic myelogenous leukemia; Cytogenetics and molecular genetics; Laboratory hematology; Lymphoproliferative disorders; Minimal residual disease