Cayuela Jean-Michel, Mauté Carole, Fabre Anne-Lise, Nibourel Olivier, Dulucq Stéphanie, Delabesse Eric, Villarèse Patrick, Hayette Sandrine, Mozziconacci Marie-Joelle, Macintyre Elizabeth
Laboratory of Hematology, University Hospital Saint-Louis, AP-HP, Paris, France; EA3518, University Paris Diderot, Paris, France.
Laboratory of Hematology, University Hospital Saint-Louis, AP-HP, Paris, France.
Clin Biochem. 2015 Oct;48(15):982-7. doi: 10.1016/j.clinbiochem.2015.04.004. Epub 2015 Apr 12.
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.
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.
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.
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.
用于分子诊断的方法的性能必须通过定期分析内部质量控制来严格控制。然而，核酸对照品的预处理、运输和长期储存仍然存在问题。因此，我们评估了由Imagene公司（法国埃夫里）开发的基于微型胶囊的创新工艺，该工艺分别用于实施设计用于淋巴细胞增殖克隆性评估和BCR-ABL1 mRNA定量的DNA和RNA对照品。
从12种细胞系中提取DNA样本，这些细胞系被选择用于与大多数BIOMED-2 PCR管进行特异性扩增。从8种表达不同BCR-ABL1转录水平的细胞系混合物中提取RNA样本。DNA和RNA由Imagene公司封装，并在室温下运送到参与的实验室。要求生物学家报告回收核酸的质量数据以及PCR结果。
封装的核酸样本很容易且高效地从微型胶囊中回收。所有实验室在DNA样本中均检测到免疫球蛋白、T细胞受体和BCL2基因座处的预期重排。各实验室之间RNA的质量一致，并且符合BCR-ABL1转录本定量所需的标准。5个实验室测得的表达水平与相应的封装前参考值在±2倍区间内。此外，模拟在室温下储存长达100年的封装RNA老化研究表明，定量结果没有偏差。
因此，Imagene微型胶囊适用于在室温下储存和分发用于基于终点或实时定量PCR的分子诊断方法能力验证的遗传物质。
