Akira Kido

A novel method for the identification of saliva by detecting oral streptococci using PCR.

We have used DNA amplification methods to detect common oral bacterial strains to test for the presence of saliva in forensic samples. Streptococcus salivarius and Streptococcus mutans were detected in various forms of saliva samples, whereas these streptococci were not detected in semen, urine, vaginal fluid, or on skin surfaces. Therefore, we demonstrated that these streptococci are promising new marker for the forensic identification of saliva. Our data indicated that S. salivarius is more reliable than S. mutans as an indicator of saliva presence, because the detection rates for S. salivarius and S. mutans by this method were 100% and 90%, respectively. Furthermore, S. salivarius was detected in all saliva stain samples, whereas S. mutans was only identified in 60% of the stains. Finally, using this method we were able to successfully detect S. salivarius and S. mutans in mock forensic samples. We therefore suggested that this method is useful for the identification of saliva in forensic science.

Typing the 1.1 kb control region of human mitochondrial DNA in Japanese individuals.

ABSTRACT: This study presents a reliable method that uses high‐fidelity long‐range PCR and optimized primers to assess polymorphism and to genotype human mitochondrial DNA (mtDNA). This method was used to analyze polymorphic sites in the human mtDNA control region, including hypervariable regions I, II, and III (HVI, HVII, and HVIII), from 124 unrelated Japanese individuals. In HVI, HVII, and HVIII, 80, 37, and 14 polymorphic sites were identified, respectively, excluding those in the homopolymeric cytosine stretch (C‐stretch) regions. The region between HVI and HVII also contained 15 polymorphic sites. On the other hand, C‐stretch length heteroplasmy in HVI or HVII was observed in 66 of 124 Japanese individuals (53%), which is much higher than in Caucasian populations. The variants in the C‐stretch regions were characterized by counting the number of heteroplasmic peaks split from the single peak in homoplasmic sequences (i.e., 16244G and 16255G in HVI and 285G in HVII). Including the C‐stretch length heteroplasmy, the 124 Japanese mtDNA samples were classified into 116 distinct haplotypes. The random match probability and the genetic diversity were estimated to be 0.95% and 0.998581, respectively, indicating that the method presented here has higher discrimination than the conventional method for mtDNA typing using HVI and HVII. [Correction added after publication 30 January 2007: in the preceding sentence random match probability and genetic diversity estimates were corrected from 0.95 and 0.998581%, respectively, to 0.95% and 0.998581, respectively.] The haplogroups and their frequencies observed in this study (i.e., D4; 13.7%, M7a1; 11.3%, D4a; 9.7% and M7b2; 8.9%) were similar to those observed in other studies of Japanese mtDNA polymorphism. The method described here is suitable for forensic applications, as shown by successful analysis of tissues from highly putrefied remains of an infant, which allowed maternal relationship to be determined via mtDNA haplotyping.

Methamphetamine body packer : Acute poisoning death due to massive leaking of methamphetamine

We encountered three methamphetamine (MA) body packers presenting simultaneously, one of whom died. Three Nigerian men (39, 35, and 37u2003years old) who attempted to smuggle were found to contain 35 (498u2003g), 21 (292u2003g), and 5 packages (73u2003g) of methamphetamine hydrochloride (MA‐HCl) in their stomachs, respectively. Packages were wrapped with plastic film and Scotch tape. The 39‐year‐old man died with acute poisoning from c. 20u2003g of MA‐HCl that had leaked from the packages into the stomach. His plasma MA concentration was 8.6u2003μg/mL when he was hospitalized (17u2003h before his death). Autopsy findings showed extreme pulmonary congestion and edema as well as moderate hepatic edema and several petechiae. Quantitative analysis was performed by gas chromatography/mass spectrometry. Extremely high concentrations of MA and its metabolite amphetamine (AP) were found in cardiac blood (63.5u2003μg/mL and 1.2u2003μg/mL), urine (4,518u2003μg/mL and 72.4u2003μg/mL), gastric contents (8,490u2003μg/mL and 16.9u2003μg/mL), and in all other autopsy samples. These high concentrations confirmed that the cause of death was acute MA poisoning. Furthermore, impurity‐profiling analysis of the seized MA revealed that the MA smuggled by the three suspects originated from the same batch.

Population genetics of 10 Y-chromosomal STR loci in Japanese using multiplexed PCR amplification kits, Y-PLEX™6 and Y-PLEX™5

Abstract Allele and haplotype frequencies of 10 Y-STR loci DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438 and DYS439 were determined in 115 unrelated Japanese males using the Y-PLEX™6 and Y-PLEX™5 PCR amplification kits. A total of 105 different haplotypes was observed, 96 of them being unique. The haplotype diversity was 0.9960 and the discrimination capacity was 0.9130.

Nine short tandem repeat loci analysis in aged semen stains using the AmpFLSTR Profiler Kit and description of a new vWA variant allele

Nine short tandem repeat (STR) loci, D3S1358, D5S818, vWA, TH01, D13S317, TPOX, FGA, D7S820 and CSF1PO, were investigated in semen stains of various ages using the AmpFLSTR Profiler Kit. The nine STR loci were typed from semen stains stored for up to 25 years with the application of 1-10 ng DNA. This system provides a useful tool in medicolegal individualization of aged semen stains. During this investigation we found a new variant allele 18.1 at the vWA locus.

Population data of nine STR loci, D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820, in Bangladeshis and Indonesians.

Allele frequencies of nine short tandem repeat (STR) loci, D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820, were determined for 127 unrelated Bangladeshi individuals and 105 unrelated Indonesian individuals using the AmpFLSTR Profiler Kit. The genotype frequency distributions of the nine STR loci were in the Hardy-Weinberg equilibrium for both populations.

Population data for 15 STR loci D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penat D, vWA, D8S1179, TPOX and FGA in Japanese

Abstract Fifteen short tandem repeat (STR) loci D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX and FGA were analyzed in 164 unrelated Japanese using the PowerPlex® 16 System kit. The genotype frequency distribution of each locus did not deviate from the Hardy–Weinberg equilibrium. Penta E was the best STR for forensic purpose. The combined power of discrimination (PD) was 0.999999999999999978.

Population genetic data on five STR loci, vWA, TH01, TPOX, F13A01 and CSF1PO, in South African blacks and South African whites.

Abstract Allele frequencies for five STR loci, vWA, TH01, TPOX, F13A01 and CSF1PO, were investigated in South African black and South African white populations. The population data did not deviate from the Hardy-Weinberg equilibrium in either population