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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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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. |
format | Online Article Text |
id | pubmed-4321704 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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