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

Biological Procedures Online

Methodology

Petroula Gerasimou¹  Andri Miltiadou¹  Jianxiang Chi²  Chryso Pierides²  Andrie Mitsidou²  Paul Costeas³ 

[1] Karaiskakio Foundation, Nicosia, Cyprus;The Center for the Study of Haematological Malignancies, Nicosia, Cyprus;The Center for the Study of Haematological Malignancies, Nicosia, Cyprus;Karaiskakio Foundation, Nicosia, Cyprus;

关键词: BCR-ABL;    cDNA synthesis;    CML;    MMR;    MRD;   

DOI  :  10.1186/s12575-015-0014-x

 received in 2014-11-07, accepted in 2015-01-03,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundThe 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).ResultsHerein, 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.ConclusionsAccurate 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.

【 授权许可】

Unknown   
© Chi et al.; licensee BioMed Central. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.