JaLCDOI 10.18926/AMO/47011
FullText URL 65_5_299.pdf
Author Itani, Miki| Yamamoto, Yuji| Doi, Yusuke| Miyaishi, Satoru|
Abstract Postmortem degradation of DNA was quantitatively estimated. Brain, liver, kidney and muscle samples were obtained from sacrificed rats left at 20℃ or 4℃. The quantity of DNA was measured by real-time PCR using a primer set for a sequence in the Rsrc 1 gene. When the quantity of amplified DNA using 10ng Human Genomic DNA was defined as 100 RFU, the quantities in the brain, liver, kidney and skeletal muscle (each 2μg of dry weight) on the day of sacrifice were 253±11, 338±22, 556±14 and 531±12 Relative Fluorescence Units (RFU), respectively (mean±S.E., n=5). The quantity of amplified DNA decreased to below 10 RFU in 1-3 weeks in the liver, kidney and skeletal muscle at 20℃, while that in the brain was more than 10 RFU for six weeks, demonstrating the usefulness of the brain as a sample for DNA analysis of decaying corpses. It was suggested that quantifying the amplified DNA in the brain at 20℃ and in the liver at 4℃ as well as the ratio of the quantity of amplified DNA in the liver to the brain at 4℃ might be useful for diagnosing time of death. This study provides the first quantitative analysis of the postmortem progress of DNA degradation in the corpse.
Keywords DNA degradation postmortem interval personal identification
Amo Type Original Article
Published Date 2011-10
Publication Title Acta Medica Okayama
Volume volume65
Issue issue5
Publisher Okayama University Medical School
Start Page 299
End Page 306
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
Copyright Holders CopyrightⒸ 2011 by Okayama University Medical School
File Version publisher
Refereed True
PubMed ID 22037266
Web of Sience KeyUT 000296116400003
JaLCDOI 10.18926/AMO/32309
FullText URL fulltext.pdf
Author Murakami, Hiroki| Ymamamoto, Yuji| Yoshitome, Kei| Ono, Toshiaki| Okamoto, Osamu| Shigeta, Yoshiaki| Doi, Yusuke| Miyaishi, Satoru| Ishizu, Hideo|
Abstract <p>In this study, sex determination using polymerase chain reaction (PCR) on tooth material was evaluated from the viewpoint of forensic medicine. The sensitivity of PCR for detection of the Y chromosome-specific alphoid repeat sequence and the X chromosome-specific alphoid repeat sequence was 0.5 pg of genomic DNA. Sex could be determined by PCR of DNA extracted from the pulp of 16 freshly extracted permanent teeth and dentine including the surface of the pulp cavity of 6 freshly extracted milk teeth. Sex could be determined using the pulp in all 20 teeth (10 male and 10 female) preserved at room temperature for 22 years. For the pulp of teeth stored in sea water, the sex could be determined in all 8 teeth immersed for 1 week and in 5 of 6 teeth immersed for 4 weeks. In the remaining 1 tooth, in which sex determination based on the pulp failed, the sex could be determined correctly when DNA extracted from the tooth hard tissue was examined. For teeth stored in soil, the sex could be determined accurately in all 8 teeth buried for 1 week, 7 of 8 teeth buried for 4 weeks, and in all 6 teeth buried for 8 weeks. When teeth were heated for 30 min, sex determination from the pulp was possible in all teeth heated to 100, 150, and 200 degrees C, and even in some teeth heated to 250 degrees C. When this method was applied to actual forensic cases, the sex of a mummified body estimated to have been discovered half a year to 1 year after death could be determined readily by examination of the dental pulp. In the skeletons of 2 bodies placed under water for approximately 1 year and approximately 11 years and 7 months, pulp tissues had been dissolved and lost, but sex determination was possible using DNA extracted from hard dental tissues. These results indicate that this method is useful in forensic practices for sex determination based on teeth samples.</p>
Keywords personal identification sex determination tooth deoxyribonucleic acid (DNA). polymerase chain reaction
Amo Type Article
Published Date 2000-02
Publication Title Acta Medica Okayama
Volume volume54
Issue issue1
Publisher Okayama University Medical School
Start Page 21
End Page 32
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 10709619
Web of Sience KeyUT 000085526000004
JaLCDOI 10.18926/AMO/31971
FullText URL fulltext.pdf
Author Imabayashi, Kiyomi| Yamamoto, Yuji| Inagaki, Sachiyo| Doi, Yusuke| Yoshitome, Kei| Miyaishi, Satoru| Ishizu, Hideo|
Abstract <p>We have improved on conventional methods for HLA-DRB1 genotyping and devised a new method that is simple, cost-effective, and adequately applicable to routine forensic practice. This method consists of group-specific polymerase chain reaction (PCR) of the exon 2 region of the HLA-DRB1 gene and simultaneous detection of single nucleotide polymorphisms (SNPs) at multiple sites using multiplex primer extension reactions. With this method, we successfully detected HLA-DRB1 genotypes from the following materials: the peripheral blood of 142 donors, 6 aged saliva stains of known DRB1 genotype stored for 5-10 years at room temperature, 10 aged bloodstains of unknown DRB1 genotype stored for 29 years at room temperature, and minimal bloodstains and saliva stains from 3 donors of known DRB1 genotypes. Furthermore, we were able to type DRB1 alleles of the minor component in mixed samples at a proportion of 1/1,000 or 1/10,000. In a criminal case, DRB1 alleles detected from mixed bloodstains on a sword found at the scene enabled us to explain the case. This method is expected to be useful for forensic medicine.</p>
Keywords HLA-DRB1 genotyping group specific primer single nucleotide polymorphism multiplex primer extension reactions application to mixed samples
Amo Type Article
Published Date 2005-10
Publication Title Acta Medica Okayama
Volume volume59
Issue issue5
Publisher Okayama University Medical School
Start Page 179
End Page 194
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 16286957
Web of Sience KeyUT 000232835600002
JaLCDOI 10.18926/AMO/31707
FullText URL fulltext.pdf
Author Shigeta, Yoshiaki| Yamamoto, Yuji| Doi, Yusuke| Miyaishi, Satoru| Ishizu, Hideo|
Abstract <p>We describe a modified method for typing a polymorphic microsatellite D12S391 locus by PCR using a newly designed primer pair. This primer pair produces shorter D12S391 amplified fragments (104-156 bp) than the primer pair originally described by Lareu et al. (209-261 bp). The detection system for the D12S391 locus using the new primer pair and capillary electrophoresis (CE) analysis was evaluated using various forensic samples. The typing results from 70 DNA samples using the new primer pair and the original primer pair were completely identical. One hundred twenty-five amplified fragments from D12S391 alleles were sized correctly within +/- 0.25 bp of the D12S391 allelic ladder. A rare allele, 19.3, previously found only in Caucasians, was found for the first time in a Japanese subject, and it was clearly distinguished from allele 20 by the CE analysis. This detection system was sensitive and could detect D12S391 types from 16 pg of genomic DNA, and from a minor component at a ratio of 1:10 in mixed samples. This system was more useful for the analysis of degraded DNA than was the method using the original primer pair, and could detect D12S391 types from bloodstains that had been stored for 26 years. In addition, the specificity of the method was demonstrated using nonhuman DNA.</p>
Keywords short tandem repeats D12S391 forensic application capillary electrophoresis
Amo Type Article
Published Date 2002-10
Publication Title Acta Medica Okayama
Volume volume56
Issue issue5
Publisher Okayama University Medical School
Start Page 229
End Page 236
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 12530506
Web of Sience KeyUT 000178668100003