Intraplatform reproducibility and technical precision of gene expression profiling in 4 laboratories investigating 160 leukemia samples: the DACH study

Clinical Chemistry
A KohlmannT Haferlach

Abstract

Gene expression profiling has the potential to offer consistent, objective diagnostic test results once a standardized protocol has been established. We investigated the robustness, precision, and reproducibility of microarray technology. One hundred sixty individual patient samples representing 11 subtypes of acute and chronic leukemias, myelodysplastic syndromes, and nonleukemia as a control group were centrally collected and diagnosed as part of the daily routine in the Munich Leukemia Laboratory. The custom AmpliChip Leukemia research microarray was used for technical analyses of quadruplicate mononuclear cell lysates in 4 different laboratories in Germany (D), Austria (A), and Switzerland (CH) (the DACH study). Total-RNA preparations were successfully performed in 637 (99.5%) of 640 cases. Mean differences between pairs of laboratories in the total-RNA yield from the same sample ranged from 0.02 mug to 1.03 mug. Further processing produced 622 successful in vitro transcription reactions (97.6%); the mean differences between laboratories in the cRNA yield from the same sample ranged from 0.40 mug to 6.18 mug. After hybridization to microarrays, a mean of 47.6%, 46.5%, 46.2%, and 46.4% of probe sets were detected as present ...Continue Reading

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Citations

Mar 30, 2011·Leukemia·K Theilgaard-MönchL Bullinger
May 29, 2009·Briefings in Functional Genomics & Proteomics·Ulrike BacherTorsten Haferlach
May 27, 2009·Current Opinion in Hematology·Krzysztof MrózekGuido Marcucci
Aug 25, 2009·Best Practice & Research. Clinical Haematology·Ulrike BacherTorsten Haferlach
Aug 25, 2009·Best Practice & Research. Clinical Haematology·Carles CodonyFrancesc Bosch
Apr 7, 2009·British Journal of Haematology·Ulrike BacherTorsten Haferlach
Jan 3, 2012·Seminars in Cell & Developmental Biology·Peter W HarrisonJudith E Mank
Apr 22, 2010·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Torsten HaferlachRobin Foà
Jun 23, 2009·Molecular Diagnosis & Therapy·Tatyana V NasedkinaAlexander S Zasedatelev

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