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Investigation into DNA recovered from human teeth for forensic applications

In South Africa, there is a burden of unidentified deceased individuals in forensic mortuaries. When human remains are severely compromised, hard tissues may provide the only DNA source for identification. The QIAamp® DNA Investigator Kit is used in forensic laboratories worldwide, including in Sout...

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Main Author: Haikney, Tarryn
Other Authors: Heathfield, Laura
Format: Thesis
Language:English
Published: Department of Clinical Laboratory Sciences 2020
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access_status_str Open Access
author Haikney, Tarryn
author2 Heathfield, Laura
author_browse Haikney, Tarryn
Heathfield, Laura
author_facet Heathfield, Laura
Haikney, Tarryn
author_sort Haikney, Tarryn
collection Thesis
description In South Africa, there is a burden of unidentified deceased individuals in forensic mortuaries. When human remains are severely compromised, hard tissues may provide the only DNA source for identification. The QIAamp® DNA Investigator Kit is used in forensic laboratories worldwide, including in South Africa, to extract DNA for identification purposes. However, in local forensic casework, the DNA recovered from teeth is often of insufficient quantity and quality for generating a DNA profile. The phenol-chloroform DNA extraction method has demonstrated improved, yet inconsistent results, when used on hard tissues. Therefore, this study assessed DNA recovery from 52 human control teeth from three deceased individuals, using an optimised phenol-chloroform method. This method involved an overnight demineralisation, two additions of phenol/chloroform/isoamyl alcohol (25:24:1) and an ethanol precipitation, as used by the Australian Federal Police. Quantitative PCR (Quantifiler™ Trio DNA Quantification Kit) and DNA profiling (PowerPlex® ESI 16 System) were then used to assess DNA quantity and quality. Results were compared to those obtained from the same teeth but extracted using the QIAamp® DNA Investigator Kit. The phenol-chloroform method recovered DNA with significantly higher yields (p = 0.0454) and significantly less degradation (p < 0.0001). Despite this improvement, there was no significant difference in DNA profiling success. This study also did a preliminary analysis of other factors affecting results and suggested that premolars might be the best tooth type with regards to DNA quantity, quality and profiling. Furthermore, dental disease and jawbone had a significant impact on results from teeth. Lastly, the phenol-chloroform method was applied to six teeth from a marine decomposition case to assess its performance in a local forensic setting. DNA metrics were particularly poor in this casework example, highlighting how different forensic and control environments are and the need for further optimisation. Overall, this study supports the use of the phenol-chloroform method and has provided a preliminary suggestion of the best tooth type, jawbone and tooth condition for DNA analysis for forensic human identification.
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institution University of Cape Town (South Africa)
language eng
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2020
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spelling oai:open.uct.ac.za:11427/32223 Investigation into DNA recovered from human teeth for forensic applications Haikney, Tarryn Heathfield, Laura Gibbon Victoria Forensic Medicine and Toxicology In South Africa, there is a burden of unidentified deceased individuals in forensic mortuaries. When human remains are severely compromised, hard tissues may provide the only DNA source for identification. The QIAamp® DNA Investigator Kit is used in forensic laboratories worldwide, including in South Africa, to extract DNA for identification purposes. However, in local forensic casework, the DNA recovered from teeth is often of insufficient quantity and quality for generating a DNA profile. The phenol-chloroform DNA extraction method has demonstrated improved, yet inconsistent results, when used on hard tissues. Therefore, this study assessed DNA recovery from 52 human control teeth from three deceased individuals, using an optimised phenol-chloroform method. This method involved an overnight demineralisation, two additions of phenol/chloroform/isoamyl alcohol (25:24:1) and an ethanol precipitation, as used by the Australian Federal Police. Quantitative PCR (Quantifiler™ Trio DNA Quantification Kit) and DNA profiling (PowerPlex® ESI 16 System) were then used to assess DNA quantity and quality. Results were compared to those obtained from the same teeth but extracted using the QIAamp® DNA Investigator Kit. The phenol-chloroform method recovered DNA with significantly higher yields (p = 0.0454) and significantly less degradation (p < 0.0001). Despite this improvement, there was no significant difference in DNA profiling success. This study also did a preliminary analysis of other factors affecting results and suggested that premolars might be the best tooth type with regards to DNA quantity, quality and profiling. Furthermore, dental disease and jawbone had a significant impact on results from teeth. Lastly, the phenol-chloroform method was applied to six teeth from a marine decomposition case to assess its performance in a local forensic setting. DNA metrics were particularly poor in this casework example, highlighting how different forensic and control environments are and the need for further optimisation. Overall, this study supports the use of the phenol-chloroform method and has provided a preliminary suggestion of the best tooth type, jawbone and tooth condition for DNA analysis for forensic human identification. 2020-09-11T12:06:05Z 2020-09-11T12:06:05Z 2020 2020-09-11T11:57:59Z Master Thesis Masters MPhil http://hdl.handle.net/11427/32223 eng application/pdf Department of Clinical Laboratory Sciences Faculty of Health Sciences
spellingShingle Forensic Medicine and Toxicology
Haikney, Tarryn
Investigation into DNA recovered from human teeth for forensic applications
thesis_degree_str Master's
title Investigation into DNA recovered from human teeth for forensic applications
title_full Investigation into DNA recovered from human teeth for forensic applications
title_fullStr Investigation into DNA recovered from human teeth for forensic applications
title_full_unstemmed Investigation into DNA recovered from human teeth for forensic applications
title_short Investigation into DNA recovered from human teeth for forensic applications
title_sort investigation into dna recovered from human teeth for forensic applications
topic Forensic Medicine and Toxicology
url http://hdl.handle.net/11427/32223
work_keys_str_mv AT haikneytarryn investigationintodnarecoveredfromhumanteethforforensicapplications