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Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study
BACKGROUND: The development of next‐generation sequencing approaches has accelerated the diagnostic process, although at present, there is a lack of a clear consensus on efficient management of human samples for downstream applications. This study aims to investigate timeframe (in terms of short pre...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279999/ https://www.ncbi.nlm.nih.gov/pubmed/35666551 http://dx.doi.org/10.1002/jcla.24531 |
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author | Mordenti, Marina Capicchioni, Valentina Corsini, Serena Locatelli, Manuela Abelli, Elisabetta Banchelli, Federico Boarini, Manila Sangiorgi, Luca |
author_facet | Mordenti, Marina Capicchioni, Valentina Corsini, Serena Locatelli, Manuela Abelli, Elisabetta Banchelli, Federico Boarini, Manila Sangiorgi, Luca |
author_sort | Mordenti, Marina |
collection | PubMed |
description | BACKGROUND: The development of next‐generation sequencing approaches has accelerated the diagnostic process, although at present, there is a lack of a clear consensus on efficient management of human samples for downstream applications. This study aims to investigate timeframe (in terms of short preservation), temperature, and additional preservation procedures (i.e., freeze and thaw cycles) for human biospecimens to implement the reliability and reproducibility of molecular investigations. METHODS: Overall, 45 whole peripheral bloods, 22 peripheral blood mononuclear cells samples, 15 saliva, and 15 buccal swab biospecimens (through the extracted DNA) were investigated, assessing yield, integrity, amplifiability, and sizing accuracy via the most common molecular techniques. RESULTS: Based on the overall evaluation criteria, the results indicate that DNA extracted from all samples, shortly preserved, have suitable quality and reliable reproducibility to be used in diagnostic activities and biomedical research, even if DNA from peripheral blood mononuclear cells is more affected by the experimental conditions. CONCLUSION: Our findings confirm the reliability of peripheral blood samples in almost all the experimental conditions. Saliva and buccal swabs are efficient almost as well, while peripheral blood mononuclear cells, albeit remain a primary source of DNA for molecular screenings, represent a less efficient source. |
format | Online Article Text |
id | pubmed-9279999 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-92799992022-07-15 Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study Mordenti, Marina Capicchioni, Valentina Corsini, Serena Locatelli, Manuela Abelli, Elisabetta Banchelli, Federico Boarini, Manila Sangiorgi, Luca J Clin Lab Anal Research Articles BACKGROUND: The development of next‐generation sequencing approaches has accelerated the diagnostic process, although at present, there is a lack of a clear consensus on efficient management of human samples for downstream applications. This study aims to investigate timeframe (in terms of short preservation), temperature, and additional preservation procedures (i.e., freeze and thaw cycles) for human biospecimens to implement the reliability and reproducibility of molecular investigations. METHODS: Overall, 45 whole peripheral bloods, 22 peripheral blood mononuclear cells samples, 15 saliva, and 15 buccal swab biospecimens (through the extracted DNA) were investigated, assessing yield, integrity, amplifiability, and sizing accuracy via the most common molecular techniques. RESULTS: Based on the overall evaluation criteria, the results indicate that DNA extracted from all samples, shortly preserved, have suitable quality and reliable reproducibility to be used in diagnostic activities and biomedical research, even if DNA from peripheral blood mononuclear cells is more affected by the experimental conditions. CONCLUSION: Our findings confirm the reliability of peripheral blood samples in almost all the experimental conditions. Saliva and buccal swabs are efficient almost as well, while peripheral blood mononuclear cells, albeit remain a primary source of DNA for molecular screenings, represent a less efficient source. John Wiley and Sons Inc. 2022-06-06 /pmc/articles/PMC9279999/ /pubmed/35666551 http://dx.doi.org/10.1002/jcla.24531 Text en © 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Research Articles Mordenti, Marina Capicchioni, Valentina Corsini, Serena Locatelli, Manuela Abelli, Elisabetta Banchelli, Federico Boarini, Manila Sangiorgi, Luca Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title | Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title_full | Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title_fullStr | Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title_full_unstemmed | Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title_short | Preanalytical DNA assessment for downstream applications: How to optimize the management of human biospecimens to support molecular diagnosis—An experimental study |
title_sort | preanalytical dna assessment for downstream applications: how to optimize the management of human biospecimens to support molecular diagnosis—an experimental study |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279999/ https://www.ncbi.nlm.nih.gov/pubmed/35666551 http://dx.doi.org/10.1002/jcla.24531 |
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