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Published in: International Journal of Legal Medicine 1/2017

01-01-2017 | Original Article

Comparison of DNA yield and STR success rates from different tissues in embalmed bodies

Authors: Amanda Wheeler, Natalia Czado, David Gangitano, Meredith Turnbough, Sheree Hughes-Stamm

Published in: International Journal of Legal Medicine | Issue 1/2017

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Abstract

Formalin fixation is commonly used to preserve tissue sections for pathological testing and embalming cadavers for medical dissection or burial. DNA extracted from formalin-fixed tissues may also provide an alternative source of genetic material for medical diagnosis and forensic casework, such as identifying unknown embalmed human remains. Formaldehyde causes DNA damage, chemical modifications, and degradation, thereby reducing the quantity and quality of DNA available for downstream genetic analyses. By comparing the DNA yield, level of DNA degradation, and short tandem repeat (STR) success of various tissue types, this study is the first of its kind to provide some guidance on which samples from embalmed bodies are likely to generate more complete STR profiles. Tissue samples were dissected from three male embalmed cadavers and included bone, cartilage, hair, muscle, internal organs, skin, teeth, and nail clippings. DNA was purified from all samples using the QIAamp® FFPE Tissue Kit (Qiagen), quantified using the QuantiFiler® Trio DNA Quantification kit (Life Technologies), and genotyped using the GlobalFiler® PCR Amplification Kit (Life Technologies). Results of this study showed variation in DNA quantity and STR success between different types of tissues and some variation between cadavers. Overall, bone marrow samples resulted in the highest DNA yields, the least DNA degradation, and greatest STR success. However, several muscle, hair, and nail samples generated higher STR success rates than traditionally harvested bone and tooth samples. A key advantage to preferentially using these tissue samples over bone (and marrow) and teeth is their comparative ease and speed of collection from the cadaver and processing during DNA extraction. Results also indicate that soft tissues affected by lividity (blood pooling) may experience greater exposure to formalin, resulting in more DNA damage and reduced downstream STR success than tissues under compression. Overall, we recommend harvesting from selected muscles (gastrocnemius, rectus femoris, flexor digitorum brevis, masseter, brachioradialis) or fingernails for human identification purposes.
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Literature
1.
go back to reference Farrugia A, Keyser C, Ludes B (2010) Efficiency evaluation of a DNA extraction and purification protocol on archival formalin-fixed and paraffin-embedded tissue. Forensic Sci Int 194:25–28CrossRef Farrugia A, Keyser C, Ludes B (2010) Efficiency evaluation of a DNA extraction and purification protocol on archival formalin-fixed and paraffin-embedded tissue. Forensic Sci Int 194:25–28CrossRef
2.
go back to reference Funabashi K et al. (2012) DNA extraction and molecular analysis of non-tumoral liver, spleen, and brain from autopsy samples: the effect of formalin fixation and paraffin embedding. Pathol Res Pract:584–591 Funabashi K et al. (2012) DNA extraction and molecular analysis of non-tumoral liver, spleen, and brain from autopsy samples: the effect of formalin fixation and paraffin embedding. Pathol Res Pract:584–591
3.
go back to reference Baden M (2005) Exhumation. In: Spitze W, Spitz D (eds) Medicolegal investigation of death: guidelines for the application of pathology to crime investigation, 4th edn. Springfield, Illinois, pp. 174–183 Baden M (2005) Exhumation. In: Spitze W, Spitz D (eds) Medicolegal investigation of death: guidelines for the application of pathology to crime investigation, 4th edn. Springfield, Illinois, pp. 174–183
5.
go back to reference Okello J. B, et al (2010) Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues. Anal Biochem 1:110–117. Okello J. B, et al (2010) Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues. Anal Biochem 1:110–117.
7.
go back to reference Hansen J et al. (2014) DNA and RNA analysis of blood and muscle from bodies with variable postmortem intervals. Forensic Sci Med Pathol 3:322–328CrossRef Hansen J et al. (2014) DNA and RNA analysis of blood and muscle from bodies with variable postmortem intervals. Forensic Sci Med Pathol 3:322–328CrossRef
8.
go back to reference Taguchi M et al. (2012) DNA identification of formalin-fixed organs is affected by fixation time and type of fixatives: using the AmpF l STR(R) Identifiler(R) PCR Amplification Kit. Med Sci Law 1:12–16CrossRef Taguchi M et al. (2012) DNA identification of formalin-fixed organs is affected by fixation time and type of fixatives: using the AmpF l STR(R) Identifiler(R) PCR Amplification Kit. Med Sci Law 1:12–16CrossRef
9.
go back to reference Lin J et al. (2009) High-quality genomic DNA extraction from formalin-fixed and paraffin-embedded samples deparaffinized using mineral oil. Anal Biochem 2:265–267CrossRef Lin J et al. (2009) High-quality genomic DNA extraction from formalin-fixed and paraffin-embedded samples deparaffinized using mineral oil. Anal Biochem 2:265–267CrossRef
10.
go back to reference Ludyga N (2012) Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses. Virchows Arch 2:131–140CrossRef Ludyga N (2012) Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses. Virchows Arch 2:131–140CrossRef
11.
go back to reference Zagga A et al. (2013) PCR inhibitory effects of aldehyde fixing agents on DNA extracted from embalmed human skeletal fragments and teeth specimens. Nurs Health Sci 1:33–37 Zagga A et al. (2013) PCR inhibitory effects of aldehyde fixing agents on DNA extracted from embalmed human skeletal fragments and teeth specimens. Nurs Health Sci 1:33–37
12.
go back to reference Korthuis R (2011) Skeletal muscle circulation. Colloquium series on integrated systems physiology. Morgan & Claypool Life Sciences, San Rafael Korthuis R (2011) Skeletal muscle circulation. Colloquium series on integrated systems physiology. Morgan & Claypool Life Sciences, San Rafael
14.
go back to reference Saladin K. (2011) Support and movement. In: Anatomy & Physiology, 6th edn. New York City, New York, pp 180–309 Saladin K. (2011) Support and movement. In: Anatomy & Physiology, 6th edn. New York City, New York, pp 180–309
15.
go back to reference Goff L (2009) Early post-mortem changes and stages of decomposition in exposed cadavers. Exp Appl Acarol 1:21–36CrossRef Goff L (2009) Early post-mortem changes and stages of decomposition in exposed cadavers. Exp Appl Acarol 1:21–36CrossRef
16.
go back to reference Mathur A (2011) An overview of methods used for estimation of time since death. Aust J Forensic Sci 4:275–285CrossRef Mathur A (2011) An overview of methods used for estimation of time since death. Aust J Forensic Sci 4:275–285CrossRef
17.
go back to reference Mundorff A, Davoren J (2014) Examination of DNA yield rates for different skeletal elements at increasing post mortem intervals. Forensic Sci Int Genet 8:55–63CrossRefPubMed Mundorff A, Davoren J (2014) Examination of DNA yield rates for different skeletal elements at increasing post mortem intervals. Forensic Sci Int Genet 8:55–63CrossRefPubMed
18.
go back to reference Pooniya S et al. (2014) Quality and quantity of extracted deoxyribonucleic acid (DNA) from preserved soft tissues of putrefied unidentifiable human corpse. J Lab Phys 6:31–35 Pooniya S et al. (2014) Quality and quantity of extracted deoxyribonucleic acid (DNA) from preserved soft tissues of putrefied unidentifiable human corpse. J Lab Phys 6:31–35
19.
go back to reference Topp T et al. (2012) Embalmed and fresh frozen human bones in orthopedic cadaveric studies: which bone is authentic and feasible? Acta Orthop 83:543–547CrossRefPubMedPubMedCentral Topp T et al. (2012) Embalmed and fresh frozen human bones in orthopedic cadaveric studies: which bone is authentic and feasible? Acta Orthop 83:543–547CrossRefPubMedPubMedCentral
22.
go back to reference LifeTechnologies (2014) GlobalFiler PCR Amplification Kit. Thermo Fisher Scientific Brand. Accessed on 27 April 2015 LifeTechnologies (2014) GlobalFiler PCR Amplification Kit. Thermo Fisher Scientific Brand. Accessed on 27 April 2015
23.
go back to reference Westen A, Gerretsen R, Maat G (2008) Femur, rib, and tooth sample collection for DNA analysis in disaster victim identification (DVI): a method to minimize contamination risk. Forensic Sci Med Pathol 4:15–21CrossRefPubMed Westen A, Gerretsen R, Maat G (2008) Femur, rib, and tooth sample collection for DNA analysis in disaster victim identification (DVI): a method to minimize contamination risk. Forensic Sci Med Pathol 4:15–21CrossRefPubMed
24.
go back to reference von Wurmb-Schwark N et al. (2008) The impact of DNA contamination of bone samples in forensic case analysis and anthropological research. Legal Med 10:125–130CrossRef von Wurmb-Schwark N et al. (2008) The impact of DNA contamination of bone samples in forensic case analysis and anthropological research. Legal Med 10:125–130CrossRef
25.
go back to reference Ossowski A et al. (2013) Example of human individual identification from World War II gravesite. Forensic Sci Int 233:179–192CrossRefPubMed Ossowski A et al. (2013) Example of human individual identification from World War II gravesite. Forensic Sci Int 233:179–192CrossRefPubMed
26.
go back to reference Vernarecci S et al. (2015) Quantifiler Trio Kit and forensic samples management: a matter of degradation. Forensic Sci Int Genet 16:77–85CrossRefPubMed Vernarecci S et al. (2015) Quantifiler Trio Kit and forensic samples management: a matter of degradation. Forensic Sci Int Genet 16:77–85CrossRefPubMed
Metadata
Title
Comparison of DNA yield and STR success rates from different tissues in embalmed bodies
Authors
Amanda Wheeler
Natalia Czado
David Gangitano
Meredith Turnbough
Sheree Hughes-Stamm
Publication date
01-01-2017
Publisher
Springer Berlin Heidelberg
Published in
International Journal of Legal Medicine / Issue 1/2017
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI
https://doi.org/10.1007/s00414-016-1405-5

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