title: cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit.
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.
authors: The Center for the Study of Haematological Malignancies, Nicosia, Cyprus.
authors: Jianxiang
authors: J
authors: Chi
authors: The Center for the Study of Haematological Malignancies, Nicosia, Cyprus.
authors: Chryso
authors: C
authors: Pierides
authors: The Center for the Study of Haematological Malignancies, Nicosia, Cyprus.
authors: Andrie
authors: A
authors: Mitsidou
authors: Karaiskakio Foundation, Nicosia, Cyprus.
authors: Andri
authors: A
authors: Miltiadou
authors: Karaiskakio Foundation, Nicosia, Cyprus.
authors: Petroula
authors: P
authors: Gerasimou
authors: The Center for the Study of Haematological Malignancies, Nicosia, Cyprus.|Karaiskakio Foundation, Nicosia, Cyprus.
authors: Paul
authors: P
authors: Costeas
keywords: BCR-ABL; CML; MMR; MRD; cDNA synthesis