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Forensic Sci Int Genet. 2014 May;10:17-22. doi: 10.1016/j.fsigen.2014.01.001. Epub 2014 Jan 13.

DNA extraction for short tandem repeat typing from mixed samples using anti-human leukocyte CD45 and ABO blood group antibodies.

Author information

  • 1Department of Legal Medicine, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
  • 2Department of Legal Medicine, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan. Electronic address: k-honda@md.tsukuba.ac.jp.
  • 3Department of Legal Medicine, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan; Faculty of Nursing, Bunri University of Hospitality, 311-1 Kashiwabara-Shinden, Sayama, Saitama 350-1336, Japan.
  • 4Department of Legal Medicine, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan; Department of Legal Medicine, Dokkyo Medical University, 880 Kitakobayashi, Mibu-machi, Shimotsuga-gun, Tochigi 321-0293, Japan.

Abstract

DNA testing from mixed cell samples can be difficult to use successfully in criminal investigations. Here, we present a method for the extraction of DNA from mixed bloodstains involving plural contributors, after antibody-microbead captured cell separation. This method, together with the multiplex short tandem repeat typing presented, has proven highly successful in the recovery of DNA profiles corresponding to the ABO blood type. Methodological steps include magnetic separation using leukocyte specific CD45 antibody-coated microbeads and centrifugal separation of leukocyte agglutination by ABO antibody. The detection results of variable mixed ratio showed that the target DNA was detected accurately as low as 1:512 mixed ratio, regardless of the large amount of the background DNA present. The method presented here is applicable to PCR-based identification for various kinds of mixed samples.

Copyright © 2014. Published by Elsevier Ireland Ltd.

KEYWORDS:

ABO antibody; CD45 microbead; DNA extraction; Mixed stain; STR analysis

PMID:
24680125
[PubMed - indexed for MEDLINE]
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