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Detecting DNA damage in stored blood samples
Several commercially available quantitative real-time PCR (qPCR) systems enable highly sensitive detection of human DNA and provide a degradation index (DI) to assess DNA quality. From routine casework in forensic genetics, it was observed that DNA degradation in forensic samples such as blood sampl...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10014702/ https://www.ncbi.nlm.nih.gov/pubmed/36355320 http://dx.doi.org/10.1007/s12024-022-00549-3 |
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author | Schulze Johann, Kristina Bauer, Hannah Wiegand, Peter Pfeiffer, Heidi Vennemann, Marielle |
author_facet | Schulze Johann, Kristina Bauer, Hannah Wiegand, Peter Pfeiffer, Heidi Vennemann, Marielle |
author_sort | Schulze Johann, Kristina |
collection | PubMed |
description | Several commercially available quantitative real-time PCR (qPCR) systems enable highly sensitive detection of human DNA and provide a degradation index (DI) to assess DNA quality. From routine casework in forensic genetics, it was observed that DNA degradation in forensic samples such as blood samples stored under sub-optimal conditions leads to visible effects in multiplex analyses of short tandem repeat markers (STRs) due to decreased amplification efficiencies in longer amplicons. It was further noticed that degradation indices often remain below the value that is considered to be critical. Thus, the aim of this work was to systematically analyze this effect and to compare conventional qPCR assays with a modified qPCR approach using uracil DNA glycosylase (UNG) and DNA quality assessment methods based on electrophoresis. Blood samples were stored at three different storage temperatures for up to 316 days. Significantly increased DNA recovery was observed from samples stored at high temperatures (37 °C) compared samples stored at room temperature and 4 °C. We observed typical effects of degradation in STR analyses but no correlation between DI and storage time in any of the storage conditions. Adding UNG slightly increased the sensitivity of detecting DNA degradation in one of the qPCR kits used in this study. This observation was not confirmed when using a second qPCR system. Electrophoretic systems did also not reveal significant correlations between integrity values and time. Methods for detecting DNA degradation are usually limited to the detection of DNA fragmentation, and we conclude that degradation affecting forensic STR typing is more complex. |
format | Online Article Text |
id | pubmed-10014702 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-100147022023-03-16 Detecting DNA damage in stored blood samples Schulze Johann, Kristina Bauer, Hannah Wiegand, Peter Pfeiffer, Heidi Vennemann, Marielle Forensic Sci Med Pathol Original Article Several commercially available quantitative real-time PCR (qPCR) systems enable highly sensitive detection of human DNA and provide a degradation index (DI) to assess DNA quality. From routine casework in forensic genetics, it was observed that DNA degradation in forensic samples such as blood samples stored under sub-optimal conditions leads to visible effects in multiplex analyses of short tandem repeat markers (STRs) due to decreased amplification efficiencies in longer amplicons. It was further noticed that degradation indices often remain below the value that is considered to be critical. Thus, the aim of this work was to systematically analyze this effect and to compare conventional qPCR assays with a modified qPCR approach using uracil DNA glycosylase (UNG) and DNA quality assessment methods based on electrophoresis. Blood samples were stored at three different storage temperatures for up to 316 days. Significantly increased DNA recovery was observed from samples stored at high temperatures (37 °C) compared samples stored at room temperature and 4 °C. We observed typical effects of degradation in STR analyses but no correlation between DI and storage time in any of the storage conditions. Adding UNG slightly increased the sensitivity of detecting DNA degradation in one of the qPCR kits used in this study. This observation was not confirmed when using a second qPCR system. Electrophoretic systems did also not reveal significant correlations between integrity values and time. Methods for detecting DNA degradation are usually limited to the detection of DNA fragmentation, and we conclude that degradation affecting forensic STR typing is more complex. Springer US 2022-11-10 2023 /pmc/articles/PMC10014702/ /pubmed/36355320 http://dx.doi.org/10.1007/s12024-022-00549-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Original Article Schulze Johann, Kristina Bauer, Hannah Wiegand, Peter Pfeiffer, Heidi Vennemann, Marielle Detecting DNA damage in stored blood samples |
title | Detecting DNA damage in stored blood samples |
title_full | Detecting DNA damage in stored blood samples |
title_fullStr | Detecting DNA damage in stored blood samples |
title_full_unstemmed | Detecting DNA damage in stored blood samples |
title_short | Detecting DNA damage in stored blood samples |
title_sort | detecting dna damage in stored blood samples |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10014702/ https://www.ncbi.nlm.nih.gov/pubmed/36355320 http://dx.doi.org/10.1007/s12024-022-00549-3 |
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