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. 2015 Feb 9;17(1):4.
doi: 10.1186/s12575-015-0014-x. eCollection 2015.

cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit

Jianxiang Chi #  1 , Chryso Pierides #  1 , Andrie Mitsidou  1 , Andri Miltiadou  2 , Petroula Gerasimou  2 , Paul Costeas  1   2
Affiliations

cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit

Jianxiang Chi et al. Biol Proced Online. .

Abstract

Background: The synthesis of complementary DNA (cDNA) for use in the detection of BCR-ABL1 at the Major Molecular Response (MMR) level is a well-established method used by clinical laboratories world-wide. However, the quality of cDNA provides sensitivity challenges and consequently affects the detection of Minimal Residual Disease (MRD).

Results: Herein, we evaluated six commercially available kits for the synthesis of cDNA according to amplification success rate, linearity and ABL1 copy number. Based on our results, the Invitrogen SuperScript® III Reverse Transcriptase kit performed better, among the ones used in this study, for the cDNA synthesis, followed by the First Strand cDNA Synthesis Kit for RT-PCR (AMV), available from Roche Applied Sciences.

Conclusions: Accurate and sensitive testing for the detection of abnormal transcripts, allows the correct stratification and treatment of patients. Hence, the use of a suitable kit for the cDNA synthesis is of great importance. This study provides a comprehensive point of reference for clinical laboratories in an attempt to optimize BCR-ABL1 detection. We propose that the Invitrogen SuperScript® III Reverse Transcriptase kit is the most suitable, among the ones used in this study, for the cDNA synthesis to be used for the detection of BCR-ABL1 at the MMR level in a CML MRD assay.

Keywords: BCR-ABL; CML; MMR; MRD; cDNA synthesis.

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