International Journal of Legal Medicine

, Volume 120, Issue 6, pp 372–376

Forensic mass screening using mtDNA

  • Reinhard Szibor
  • Ines Plate
  • Herrmann Schmitter
  • Holger Wittig
  • Dieter Krause
Case Report


At the forensic autopsy of a sexual murder victim, some trace hairs, possibly belonging to the perpetrator, were saved. Initially, the analysis of a pubic hair shaft only revealed the presence of the mitochondrial (mt) DNA haplotype profile consisting of the (CA)6 allele and the complete hypervariable region 1 (HV1) and 2 (HV2) sequence. Later, typing of some further telogene trace hairs, which had been stored for several years, yielded a nuclear short tandem repeat (STR) profile. We used both the mtDNA haplotype and the STR profile to start a DNA mass screening project involving 2,335 male citizens of the relevant communities. MtDNA screening was carried out by using the CA repeat amplification in combination with an SNP typing procedure based on the restriction site analysis of amplified d-loop sequences. The aim of our paper is to put mass screening with mtDNA up for discussion.


MtDNA Mass screening Dinucleotide repeat Sexual murder Restriction sites 


  1. 1.
    Anderson S, Bankier AT, Barrell BG et al (1981) Sequence and organization of the human mitochondrial genome. Nature 290:457–465CrossRefPubMedGoogle Scholar
  2. 2.
    Benedictis G de, Rose G, Passarino G et al (1989) Restriction fragment length polymorphism of human mitochondrial DNA in a sample population from Apulia (southern Italy). Ann Hum Genet 53:311–318PubMedGoogle Scholar
  3. 3.
    Brandstatter A, Parsons TJ, Parson W (2003) Rapid screening of mtDNA coding region SNPs for the identification of west European Caucasian haplogroups. Int J Legal Med 117:291–298CrossRefPubMedGoogle Scholar
  4. 4.
    Budowle B, Polanskey D (2005) FBI mtDNA database: a cogent perspective. Science 307:845–847CrossRefPubMedGoogle Scholar
  5. 5.
    Budowle B, Chakraborty R, Giusti AM et al (1991) Analysis of the VNTR locus D1S80 by the PCR followed by high-resolution PAGE. Am J Hum Genet 48:137–144PubMedGoogle Scholar
  6. 6.
    Budowle B, Gyllensten U, Chakraborty R, Allen M (2005) Forensic analysis of the mitochondrial coding region and association to disease. Int J Legal Med 119:314–315CrossRefPubMedGoogle Scholar
  7. 7.
    Butler JM, Wilson MR, Reeder DJ (1998) Rapid mitochondrial DNA typing using restriction enzyme digestion of polymerase chain reaction amplicons followed by capillary electrophoresis separation with laser-induced fluorescence detection. Electrophoresis 19:119–124CrossRefPubMedGoogle Scholar
  8. 8.
    Coble MD, Just RS, O’Callaghan JE et al (2004) Single nucleotide polymorphisms over the entire mtDNA genome that increase the power of forensic testing in Caucasians. Int J Legal Med 118:137–146CrossRefPubMedGoogle Scholar
  9. 9.
    Davies KE, Speer A, Herrmann F et al (1985) Human X chromosome markers and Duchenne muscular dystrophy. Nucleic Acids Res 13:3419–3426PubMedGoogle Scholar
  10. 10.
    Divne AM, Allen M (2005) A DNA microarray system for forensic SNP analysis. Forensic Sci Int 154:111–121CrossRefPubMedGoogle Scholar
  11. 11.
    Divne AM, Nilsson M, Calloway C et al (2005) Forensic casework analysis using the HVI/HVII mtDNA linear array assay. J Forensic Sci 50:548–554CrossRefPubMedGoogle Scholar
  12. 12.
    Hellmann A, Rohleder U, Schmitter H, Wittig M (2001) STR typing of human telogen hairs—a new approach. Int J Legal Med 114:269–273PubMedCrossRefGoogle Scholar
  13. 13.
    Holland M, Parsons T (1999) Mitochondrial DNA sequence analysis—validation and use for forensic casework. Forensic Sci Rev 11:21–50Google Scholar
  14. 14.
    Horai S, Hayasaka K (1990) Intraspecific nucleotide sequence differences in the major noncoding region of human mitochondrial DNA. Am J Hum Genet 46:828–842PubMedGoogle Scholar
  15. 15.
    Krenke BE, Tereba A, Anderson SJ et al (2002) Validation of a 16-locus fluorescent multiplex system. J Forensic Sci 47:773–785PubMedGoogle Scholar
  16. 16.
    Lutz S, Weisser HJ, Heizmann J, Pollak S (1996) mtDNA as a tool for identification of human remains. Identification using mtDNA. Int J Legal Med 109:205–209CrossRefPubMedGoogle Scholar
  17. 17.
    Lutz S, Weisser HJ, Heizmann J, Pollak S (1997) A third hypervariable region in the human mitochondrial D-loop. Hum Genet 101:384PubMedGoogle Scholar
  18. 18.
    Lutz S, Weisser HJ, Heizmann J, Pollak S (1998) Location and frequency of polymorphic positions in the mtDNA control region of individuals from Germany. Int J Legal Med 111:67–77CrossRefPubMedGoogle Scholar
  19. 19.
    Lutz S, Wittig H, Weisser HJ et al (2000) Is it possible to differentiate mtDNA by means of HVIII in samples that cannot be distinguished by sequencing the HVI and HVII regions? Forensic Sci Int 113:97–101CrossRefPubMedGoogle Scholar
  20. 20.
    Lutz-Bonengel S, Schmidt U, Schmitt T, Pollak S (2003) Sequence polymorphisms within the human mitochondrial genes MTATP6, MTATP8 and MTND4. Int J Legal Med 117:133–142PubMedGoogle Scholar
  21. 21.
    Monson K, Miller K, Wilson M, DiZinno J, Budowle B (2002) The mtDNA population database: an integrated software and database resource for forensic comparison. Forensic Sci Commun 4(2) (
  22. 22.
    Morley JM, Bark JE, Evans CE, Perry JG, Hewitt CA, Tully G (1999) Validation of mitochondrial DNA minisequencing for forensic casework. Int J Legal Med 112:241–248CrossRefPubMedGoogle Scholar
  23. 23.
    Neuhuber F, Radacher M, Krasa B (1996) F13B and CD4 allele frequencies in an Austrian population sample. Int J Legal Med 108:227–228CrossRefPubMedGoogle Scholar
  24. 24.
    Niederstatter H, Coble MD, Grubwieser P, Parsons TJ, Parson W (2006) Characterization of mtDNA SNP typing and mixture ratio assessment with simultaneous real-time PCR quantification of both allelic states. Int J Legal Med 120(1):18–23CrossRefPubMedGoogle Scholar
  25. 25.
    Parson W, Parsons TJ, Scheithauer R, Holland MM (1998) Population data for 101 Austrian Caucasian mitochondrial DNA d-loop sequences: application of mtDNA sequence analysis to a forensic case. Int J Legal Med 111:124–132CrossRefPubMedGoogle Scholar
  26. 26.
    Parson W, Brandstatter A, Alonso A et al (2004) The EDNAP mitochondrial DNA population database (EMPOP) collaborative exercises: organisation, results and perspectives. Forensic Sci Int 139:215–226CrossRefPubMedGoogle Scholar
  27. 27.
    Pfeiffer H, Brinkmann B, Huhne J et al (1999) Expanding the forensic German mitochondrial DNA control region database: genetic diversity as a function of sample size and microgeography. Int J Legal Med 112:291–298CrossRefPubMedGoogle Scholar
  28. 28.
    Pfeiffer H, Huhne J, Ortmann C, Waterkamp K, Brinkmann B (1999) Mitochondrial DNA typing from human axillary, pubic and head hair shafts—success rates and sequence comparisons. Int J Legal Med 112:287–290CrossRefPubMedGoogle Scholar
  29. 29.
    Piercy R, Sullivan KM, Benson N, Gill P (1993) The application of mitochondrial DNA typing to the study of white Caucasian genetic identification. Int J Legal Med 106:85–90CrossRefPubMedGoogle Scholar
  30. 30.
    Quintans B, Alvarez-Iglesias V, Salas A et al. (2004) Typing of mitochondrial DNA coding region SNPs of forensic and anthropological interest using SNaPshot minisequencing. Forensic Sci Int 140:251–257CrossRefPubMedGoogle Scholar
  31. 31.
    Rolf B, Waterkamp K, Huhne J (1998) Allele frequency data for the FGA locus in eight populations. Int J Legal Med 111:55–56CrossRefPubMedGoogle Scholar
  32. 32.
    Szibor R, Plate I, Krause D, Michael M (1997) Anderson sequence matching/victim sequence mismatching primers enabled the record of proper mitochondria D-loop sequences of the components of a mixed stain. In: ISFH Symposium Program Committee (ed) Advances in research on DNA polymorphisms. Toyoshoten, TokyoGoogle Scholar
  33. 33.
    Szibor R, Michael M, Spitsyn VA et al (1997) Mitochondrial D-loop 3′ (CA)n repeat polymorphism: optimization of analysis and population data. Electrophoresis 18:2857–2860CrossRefPubMedGoogle Scholar
  34. 34.
    Szibor R, Michael M, Plate I, Krause D (2000) Efficiency of forensic mtDNA analysis. Case examples demonstrating the identification of traces. Forensic Sci Int 113:71–78CrossRefPubMedGoogle Scholar
  35. 35.
    Szibor R, Michael M, Plate I, Wittig H, Krause D (2003) Identification of the minor component of a mixed stain by using mismatch primer-induced restriction sites in amplified mtDNA. Int J Legal Med 117:160–164PubMedGoogle Scholar
  36. 36.
    Szibor U, Schneider-Stock R, Augustin C et al (2003) Variability of the mitochondrial loci nt00073 and 16519 in populations of Germany, Syria, Cameroon, Japan and Peru—a study using RFLP and Light Cycler technique. In: Brinkmann B, Carracedo A (eds) Progress in forensic genetics 9. International Congress Series 1239, pp 547–551Google Scholar
  37. 37.
    Wiegand P, Budowle B, Rand S, Brinkmann B (1993) Forensic validation of the STR systems SE 33 and TC 11. Int J Legal Med 105:315–320CrossRefPubMedGoogle Scholar
  38. 38.
    Wittig H, Augustin C, Baasner A et al (2000) Mitochondrial DNA in the central European population. Human identification with the help of the forensic mt-DNA D-loop-base database. Forensic Sci Int 113:113–118CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Reinhard Szibor
    • 1
  • Ines Plate
    • 1
  • Herrmann Schmitter
    • 2
  • Holger Wittig
    • 1
  • Dieter Krause
    • 1
  1. 1.Institut für RechtsmedizinOtto-von-Guericke-Universität MagdeburgMagdeburgGermany
  2. 2.BundeskriminalamtKriminaltechnisches InstitutWiesbadenGermany

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