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

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 dete...

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Autores principales: Chi, Jianxiang, Pierides, Chryso, Mitsidou, Andrie, Miltiadou, Andri, Gerasimou, Petroula, Costeas, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321704/
https://www.ncbi.nlm.nih.gov/pubmed/25667568
http://dx.doi.org/10.1186/s12575-015-0014-x
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author Chi, Jianxiang
Pierides, Chryso
Mitsidou, Andrie
Miltiadou, Andri
Gerasimou, Petroula
Costeas, Paul
author_facet Chi, Jianxiang
Pierides, Chryso
Mitsidou, Andrie
Miltiadou, Andri
Gerasimou, Petroula
Costeas, Paul
author_sort Chi, Jianxiang
collection PubMed
description 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12575-015-0014-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-43217042015-02-10 cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit Chi, Jianxiang Pierides, Chryso Mitsidou, Andrie Miltiadou, Andri Gerasimou, Petroula Costeas, Paul Biol Proced Online Methodology 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12575-015-0014-x) contains supplementary material, which is available to authorized users. BioMed Central 2015-02-09 /pmc/articles/PMC4321704/ /pubmed/25667568 http://dx.doi.org/10.1186/s12575-015-0014-x Text en © 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.
spellingShingle Methodology
Chi, Jianxiang
Pierides, Chryso
Mitsidou, Andrie
Miltiadou, Andri
Gerasimou, Petroula
Costeas, Paul
cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title_full cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title_fullStr cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title_full_unstemmed cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title_short cDNA synthesis for BCR-ABL1 detection at the MMR level: the importance of using the appropriate kit
title_sort cdna synthesis for bcr-abl1 detection at the mmr level: the importance of using the appropriate kit
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4321704/
https://www.ncbi.nlm.nih.gov/pubmed/25667568
http://dx.doi.org/10.1186/s12575-015-0014-x
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