Highlights
- •Hair mtDNA quantity and CR variation is extremely heterogeneous between individuals.
- •Hairs of one donor/fragments of one hair can yield slightly different CR sequences.
- •Current forensic interpretation guidelines are rarely violated.
- •Affected nps are individual and rare or hypervariable also in other tissues.
- •No hair specific mtDNA variation patterns have been detected.
Abstract
Hairs from the same donor have been found to differ in mtDNA sequence within and among
themselves and from other tissues, which impacts interpretation of results obtained
in a forensic setting. However, little is known on the magnitude of this phenomenon
and published data on systematic studies are scarce. We addressed this issue by generating
mtDNA control region (CR) profiles of >450 hair fragments from 21 donors by Sanger-type
sequencing (STS). To mirror forensic scenarios, we compared hair haplotypes from the
same donors to each other, to the corresponding buccal swab reference haplotypes and
analyzed several fragments of individual hairs. We also investigated the effects of
hair color, donor sex and age, mtDNA haplogroup and chemical treatment on mtDNA quantity,
amplification success and variation. We observed a wide range of individual CR sequence
variation. The reference haplotype was the only or most common (≥75%) hair haplotype
for most donors. However, in two individuals, the reference haplotype was only found
in about a third of the investigated hairs, mainly due to differences at highly variable
positions. Similarly, most hairs revealed the reference haplotype along their entire
length, however, about a fifth of the hairs contained up to 71% of segments with deviant
haplotypes, independent of the longitudinal position. Variation affected numerous
positions, typically restricted to the individual hair and in most cases heteroplasmic,
but also fixed (i.e. homoplasmic) substitutions were observed. While existing forensic
mtDNA interpretation guidelines were found still sufficient for all comparisons to
reference haplotypes, some comparisons between hairs from the same donor could yield
false exclusions when those guidelines are strictly followed. This study pinpoints
the special care required when interpreting mtDNA results from hair in forensic casework.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Forensic Science International: GeneticsAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Evaluation of the human hair root for DNA typing subsequent to microscopic comparison.J. Forensic Sci. 1998; 43: 305-314
- Forensic mitochondrial DNA analysis: two years of commercial casework experience in the United States.Croat. Med. J. 2001; 42: 298-303
- Mitochondrial DNA sequencing of shed hairs and saliva on robbery caps: sensitivity and matching probabilities.J. Forensic Sci. 1998; 43: 453-464
- Results of the 1999–2000 collaborative exercise and proficiency testing program on mitochondrial DNA of the GEP-ISFG: an inter-laboratory study of the observed variability in the heteroplasmy level of hair from the same donor.Forensic Sci. Int. 2002; 125: 1-7
- Sequencing of mtDNA in shed hairs: a retrospective analysis of material from forensic cases and a pre-screening method.Open Forensic Sci. J. 2012; 5: 13-22
- A single difference in mtDNA control region sequence observed between hair shaft and reference samples from a single donor.in: Proceedings from the 7th International Symposium on Human Identification 1996, Promega Corporation, Madison, WI, USA1997: 126-130
- Human hair histogenesis for the mitochondrial DNA forensic scientist.J. Forensic Sci. 2001; 46: 844-853
- The current status of microscopical hair comparisons.Sci. World J. 2001; 1: 868-878
- Forensic mitochondrial DNA analysis of 691 casework hairs.J. Forensic Sci. 2005; 50: 73-80
- In situ labeling of DNA reveals interindividual variation in nuclear DNA breakdown in hair and may be useful to predict success of forensic genotyping of hair.Int. J. Legal Med. 2012; 126: 63-70
- DNA from keratinous tissue. Part I: hair and nail.Ann. Anat. 2012; 194: 17-25
- DNA typing from single hairs.Nature. 1988; 332: 543-546
- Pitfalls in the analysis of mitochondrial DNA from ancient specimens and the consequences for forensic DNA analysis: the historical case of the putative heart of Louis XVII.Int. J. Legal Med. 2001; 115: 135-141
- Degeneration of nuclei and mitochondria in human hairs.J. Forensic Sci. 2009; 54: 346-349
- A fluorescent microscopy-screening test for efficient STR-typing of telogen hair roots.Forensic Sci. Int.-Genet. 2008; 3: 27-31
- Short tandem repeat (STR) genotyping of keratinised hair. Part 2. An optimised genomic DNA extraction procedure reveals donor dependence of STR profiles.Forensic Sci. Int. 2005; 153: 247-259
- STR typing of ancient DNA extracted from hair shafts of Siberian mummies.Forensic Sci. Int. 2007; 166: 218-229
- Improved STR typing of telogen hair root and hair shaft DNA.Electrophoresis. 2007; 28: 2835-2842
- Mitochondrial DNA typing from human axillary, pubic and head hair shafts–success rates and sequence comparisons.Int. J. Legal Med. 1999; 112: 287-290
- Mitochondrial DNA heteroplasmy.Forensic Sci. Rev. 2004; 16: 1-20
- History in a single hair.EMBO Rep. 2010; 11: 427-430
- The 2000–2001 GEP–ISFG collaborative exercise on mtDNA: assessing the cause of unsuccessful mtDNA PCR amplification of hair shaft samples.Forensic Sci. Int. 2003; 134: 46-53
- Mitochondrial DNA analysis of 114 hairs measuring less than 1 cm from a 19-year-old homicide.Invest. Genet. 2012; 3: 12
- qPCR and mtDNA SNP analysis of experimentally degraded hair samples and its application in forensic casework.Int. J. Legal Med. 2010; 124: 337-342
- Ancient mitochondrial DNA from hair.Curr. Biol. 2004; 14: R463-464
- Mini-midi-mito: adapting the amplification and sequencing strategy of mtDNA to the degradation state of crime scene samples.Forensic Sci. Int. Genet. 2009; 3: 149-153
- Histological correlates of post mortem mitochondrial DNA damage in degraded hair.Forensic Sci. Int. 2006; 156: 201-207
- Resistance of degraded hair shafts to contaminant DNA.Forensic Sci. Int. 2006; 156: 208-212
- Mitochondrial-DNA sequences in single hairs from a southern african population.Proc. Natl. Acad. Sci. U. S. A. 1989; 86: 9350-9354
- Routine use of hair root or buccal swab specimens for pcr analysis–advantages over using blood.Clin. Chim. Acta. 1992; 207: 169-174
- Strategies for automated sequencing of human mitochondrial-DNA directly from PCR products.Biotechniques. 1992; 13: 82-92
- Validation of mitochondrial DNA sequencing for forensic casework analysis.Int. J. Legal Med. 1995; 108: 68-74
- Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts.Biotechniques. 1995; 18: 662-669
- Genetic analysis of single hair shafts by automated sequence analysis of the mitochondrial D-loop region.Advances in Forensic Haemogenetics. 6. Springer, Berlin1995: 17-19
- Population data for 101 Austrian Caucasian mitochondrial DNA D-loop sequences: application of mtDNA sequence analysis to a forensic case.Int. J. Legal Med. 1998; 111: 124-132
- Mitochondrial DNA: State of Tennessee v. Paul Ware.Profiles in DNA. 1998; 1: 6-7
- Rapid and noninvasive screening of patients with mitochondrial myopathy.Hum. Mutat. 1994; 4: 132-135
- Segregation patterns of a novel mutation in the mitochondrial transfer-RNA glutamic-acid gene associated with myopathy and diabetes-mellitus.Am. J. Hum. Genet. 1995; 56: 1017-1025
- Heteroplasmy in the control region of human mitochondrial DNA.Genome Res. 1995; 5: 89-90
- Results of a collaborative study of the EDNAP group regarding mitochondrial DNA heteroplasmy and segregation in hair shafts.Forensic Sci. Int. 2004; 140: 1-11
- Variable levels of a heteroplasmic point mutation in individual hair roots.Am. J. Hum. Genet. 1997; 61: 1303-1308
- Heteroplasmy in hair: study of mitochondrial DNA third hypervariable region in hair and blood samples.J. Forensic Sci. 2010; 55: 715-718
- Characterization of mitochondrial DNA sequence heteroplasmy in blood tissue and hair as a function of hair morphology.J. Forensic Sci. 2011; 56: 46-60
- Individual human hair mitochondrial DNA control region heteroplasmy proportions in mothers and children.Mitochondrion. 2006; 6: 37-42
- Inter- and intragenerational transmission of a human mitochondrial DNA heteroplasmy among 13 maternally-related individuals and differences between and within tissues in two family members.Mitochondrion. 2003; 2: 401-414
- Heteroplasmy in hair: differences among hair and blood from the same individuals are still a matter of debate.Forensic Sci. Int. 2007; 173: 117-121
- Mitochondrial DNA heteroplasmy or artefacts-a matter of the amplification strategy?.Int. J. Legal Med. 2003; 117: 180-184
- Evidence for frequent and tissue-specific sequence heteroplasmy in human mitochondrial DNA.Mitochondrion. 2015; 20: 82-94
- Mitochondrial DNA heteroplasmy among hairs from single individuals.J. Forensic Sci. 2004; 49: 986-991
- Massively parallel sequencing of complete mitochondrial genomes from hair shaft samples.Forensic Sci. Int. Genet. 2015; 15: 8-15
- Heteroplasmy in mtDNA and the weight of evidence in forensic mtDNA analysis: a case report.Int. J. Legal Med. 2001; 114: 186-190
- A family exhibiting heteroplasmy in the human mitochondrial DNA control region reveals both somatic mosaicism and pronounced segregation of mitotypes.Hum. Genet. 1997; 100: 167-171
- Mitochondrial DNA control region diversity in hairs and body fluids of monozygotic triplets.Int. J. Legal Med. 2004; 118: 71-74
- Die Rolle der humanen mitochondrialen DNA in der Haaranalytik.in: Madea B. Mußhoff F. Haaranalytik: Technik und Interpretation in Medizin und Strafrecht. Deutscher Ärzte-Verlag, Köln2004: 364-379
- EMPOP-a forensic mtDNA database.Forensic Sci. Int. Genet. 2007; 1 (http://empop.online): 88-92
- Correlation of microscopic and mitochondrial DNA hair comparisons.J. Forensic Sci. 2002; 47: 964-967
- Strengthening Forensic Science in the United States: A Path Forward.The National Academies Press, Washington, DC2009
FBI, Press release Apr 20, 2015 last accessed on July 18, 2016 at http://www.fbi.gov/news/pressrel/press-releases/fbi-testimony-on-microscopic-hair-analysis-contained-errors-in-at-least-90-percent-of-cases-in-ongoing-review.
- Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material.Biotechniques. 1991; 10: 506-513
- A modular real-time PCR concept for determining the quantity and quality of human nuclear and mitochondrial DNA.Forensic Sci. Int. Genet. 2007; 1: 29-34
- Extended guidelines for mtDNA typing of population data in forensic science.Forensic Sci. Int. Genet. 2007; 1: 13-19
- Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA.Nat. Genet. 1999; 23: 147
- Updated comprehensive phylogenetic tree of global human mitochondrial DNA variation.Hum. Mutat. 2009; 30: E386-394
- Concept for estimating mitochondrial DNA haplogroups using a maximum likelihood approach (EMMA).Forensic Sci. Int.-Genet. 2013; 7: 601-609
- Nuclear and mitochondrial DNA quantification of various forensic materials.Forensic Sci. Int. 2006; 164: 56-64
- MtDNA sequence analysis using capillary electrophoresis and its application to the analysis of mtDNA in hair.Jpn. J. Sci. Technol. Ident. 2003; 7: 123-130
- Mitochondrial DNA in human hair shafts-existence of intra-individual differences?.Int. J. Legal Med. 1999; 112: 172-175
- Untersuchungen zu möglichen Mechanismen einer PCR-Hemmung in Haarextrakten durch Melanine.Rechtsmedizin. 2001; 11: 42-45
- Specific melanin content in human hairs and mitochondrial DNA typing success.Am. J. Forensic Med. Pathol. 2009; 30: 162-166
- Mitochondrial DNA amplification success rate as a function of hair morphology.J. Forensic Sci. 2007; 52: 40-47
- Considerations by the European DNA profiling (EDNAP) group on the working practices, nomenclature and interpretation of mitochondrial DNA profiles.Forensic Sci. Int. 2001; 124: 83-91
- DNA Commission of the International Society for Forensic Genetics: revised and extended guidelines for mitochondrial DNA typing.Forensic Sci. Int. Genet. 2014; 13: 134-142
- No relationship found between point heteroplasmy in mitochondrial DNA control region and age range, sex and haplogroup in human hairs.Mol. Biol. Rep. 2011; 38: 1219-1223
- High levels of mitochondrial DNA heteroplasmy in single hair roots: reanalysis and revision.Electrophoresis. 2003; 24: 1159-1165
- Evaluating length heteroplasmy in the human mitochondrial DNA control region.Int. J. Legal Med. 2010; 124: 133-142
- Investigation of heteroplasmy in the human mitochondrial DNA control region: a synthesis of observations from more than 5000 global population samples.J. Mol. Evol. 2009; 68: 516-527
- Detecting rare triple heteroplasmic substitutions in the mitochondrial DNA control region: a potential concern for forensic DNA studies.Cell J. 2011; 13: 103-106
- The frequency of heteroplasmy in the HVII region of mtDNA differs across tissue types and increases with age.Am. J. Hum. Genet. 2000; 66: 1384-1397
- A simplified method for mitochondrial DNA extraction from head hair shafts.J. Forensic Sci. 2005; 50: 1119-1122
Scientific Working Group on DNA Analysis Methods (SWGDAM) Interpretation guidelines for mitochondrial DNA (mtDNA) analysis for forensic DNA testing laboratories, 2013.
- Interpretation Guidelines for Mitochondrial DNA Sequencing, Proceedings from the 10th International Symposium on Human Identification 1999.Promega Corporation, Madison, WI, USA2000
- Mitochondrial DNA sequence analysis–validation and use for forensic casework.Forensic Sci. Rev. 1999; 11: 21-50
- Segregation pattern and biochemical effect of the G3460A mtDNA mutation in 27 members of LHON family.J. Neurol. Sci. 2004; 223: 149-155
- The EDNAP mitochondrial DNA population database (EMPOP) collaborative exercises: organisation, results and perspectives.Forensic Sci. Int. 2004; 139: 215-226
- Factors affecting the detection and quantification of mitochondrial point heteroplasmy using Sanger sequencing and SNaPshot minisequencing.Int. J. Legal Med. 2011; 125: 427-436
- A sensitive denaturing gradient-gel electrophoresis assay reveals a high frequency of heteroplasmy in hypervariable region 1 of the human mtDNA control region.Am. J. Hum. Genet. 2000; 67: 432-443
- Mitochondrial DNA heteroplasmy in the emerging field of massively parallel sequencing.Forensic Sci. Int. Genet. 2015; 18: 131-139
- Evaluation of next generation mtGenome sequencing using the Ion torrent personal genome machine (PGM).Forensic Sci. Int. Genet. 2013; 7: 543-549
- Different methods to determine length heteroplasmy within the mitochondrial control region.Int. J. Legal Med. 2004; 118: 274-281
- Evaluating sequence-derived mtDNA length heteroplasmy by amplicon size analysis.Forensic Sci. Int.-Gen. 2011; 5: 142-145
Article info
Publication history
Published online: July 25, 2016
Accepted:
July 23,
2016
Received in revised form:
July 20,
2016
Received:
May 29,
2016
Identification
Copyright
© 2016 Elsevier Ireland Ltd. All rights reserved.
