This report describes a non-PCR-based ABO genotyping method using Loop-Mediated Isothermal Amplification (LAMP). LAMP method, based on isothermal DNA amplification, enables the amplification of high efficiency and within a shorter time. Three single nucleotide polymorphisms (SNPs) in the ABO gene (261, 796, and 803 np) were examined using four primer sets (O, non-O, B, and non-B). Then ABO genotypes were determined from combinations of positive reactions. All procedures from amplification to detection took only 1 hour because neither DNA extraction for amplification nor electrophoresis for allele discrimination was required. These results show that ABO genotyping using LAMP is convenient and useful for routine work to support serological typing.

View full abstract

  At a fire scene caused by spontaneous combustion of drying oils and semi-drying oils, long heat storage is thought to produce thermal degraded oils in the vicinity of the fire occurrence point. In other words, even if the cause of a fire scene is difficult to judge from the appearance of the damaged scene where spontaneous combustion occurred, the possibility of spontaneous combustion might be considered more clearly by proving the existence of thermal degraded parts in oily debris. Results derived from both appearance and evidence are forensically reliable.
  It is considered that not only are changes of fatty acid composition observed in thermal degraded parts, but also changes of physical and thermodynamic properties.
  This paper focuses on thermal analyses as the method to distinguish these changes. The purpose of this paper is also to discuss the validity of thermal analyses as the method to prove the existence of thermal degraded parts in oily debris.
  Differential scanning calorimetry and thermogravimetry were applied to thermal degraded samples produced in experiments, and so the freezing point and thermal decomposition temperature were measured. Furthermore, a kinetic study was carried out based on the isoconversion method; using the value of activation energy derived from the line gradients, master curves were drawn. In the course of considering the freezing point, thermal decomposition temperature, the parallel relation among the lines and convergent state of master curve, and the distinguishable possibility of thermal degraded samples were discussed.
  In conclusion, this paper clarified that a kinetic study based on the isoconversion method allowed us to distinguish parts of a slight thermal degradation.
  This method is effective in proving thermal degraded parts existing in oily debris.

View full abstract

  A variety of evidence samples have been brought into the Identification section of the police for detection of latent fingerprints. Fumes of cyanoacrylate ester are generally used to detect latent fingerprints on flat surfaces, such as cutlery, in a case of murder. White fingerprint ridges fixed by the fumes are generally stained with dye powder or dye solution.
  We developed new reagents which can detect latent fingerprints the same method as using both the fumes of cyanoacrylate ester and fluorescent staining. First, polycyanoacrylate ester powder was produced from its monomer and water as an initiator. Then, 5 percent of p-dimethylaminocinnamaldehyde (DMAC) or p-dimethylaminobenzaldehyde (DMAB), each of which has volatility and fluorescence that could be excited by ultraviolet light or blue light respectively, was added to the polycyanoacrylate ester powder. The developed reagent depolymerized and yielded monomer at a lower temperature than polycyanoacrylate powder that polymerized in methanol containing 1% water. The mixed DMAC or DMAB can simultaneously stain and give fluorescence to the white fingerprint ridge. Added to that, exposing samples to acetic acid vapor for a short time has increased the fluorescence intensity and shifted excitation wavelength to a longer one such as ultraviolet to blue and blue to green.
  By use of this reagent, the fumes of cyanoacrylate ester and fluorescent staining was achieved simultaneously. The gaseous phase detections are specially recommended for evidences that are composed of material that is easily destroyed due to its physical contact with powder or liquid in the dyeing process.

View full abstract

  For the identification of human bloodstains, OC-Hemocatch (HC), one of the fecal occult blood test kits is generally used to obtain rapid result. However, HC showed positive reaction for some of wild animal bloodstains. In this paper, a detailed comparison of the specificity of six commercial fecal occult blood test kits has been made. As a result, two, i.e. HemoGold50 (HG) and QuickGold Hem (QG), of six kits showed both good specificity and good sensitivity. The remaining fecal occult blood test kits showed positive reaction in wild animal bloodstains, such as mustelid blood or low sensitivity. HG is simple to use and easy to store for forensic case works. Therefore, HG was considered to be the most suitable fecal occult blood test kit for human identification of blood or bloodstains.

View full abstract

  A rapid screening method for simultaneous determination of phosphorus-containing amino acid type herbicides such as glyphosate (GLYP), glufosinate (GLUF), bialaphos (BIAL) and their major metabolites, AMPA and MPPA in human samples using N-Acetyl,O-Methyl derivatives with high-performance liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) was developed. Serum sample with deporoteinization with 10% trifluoroacetic acid and urine sample without pretreatment was evaporated, and then the drid analytes were directly derivatized by trimethyl orthoacetate and acetic acid at 100°C for 30 minutes. LC analyses were perfomed on a reverse phase column (Symmetry C8) in the gradient solvent system of 5 mM ammonium formate and acetonitrile.
  Calibration curves of GLYP, GLUF, BIAL, AMPA and MPPA showed good linearity in the range of 0.1-5, 0.1-5, 0.2-5, 0.2-5 and 0.1-5 μg/ml in serum, and 0.1-5, 0.1-5, 0.5-5, 0.5-5 and 0.1-5 μg/ml in urine, respectively. The detection limits of GLYP, GLUF, BIAL, AMPA and MPPA in the selected ion monitoring (SIM) mode were 0.05, 0.03, 0.1, 0.2 and 0.05 μg/ml in serum, and were 0.05, 0.05, 0.4, 0.2 and 0.05 μg/ml in urine, respectively.

View full abstract

  Biogasoline, which contains 7 vol% of bio-ETBE (ethyl t-butyl ether), began circulating in the Metropolitan area of Japan on April 27, 2007. Among 44,000 gas stations in Japan, biogasoline is currently sold only in 100 gas stations. Name and location of these 100 gas stations are disclosed. If biogasoline is truly sold only in these gas stations, tracking a gasoline sample back to its retailer will be possible; when ETBE is detected in a gasoline sample, it is certain that the sample was sold in one of the 100 gas stations. Biogasoline was collected in the end of June, September, December 2007 and March 2008 from gas stations of 3 different brands in Chiba prefecture, and its color, density, composition and concentration of ETBE were analyzed. Regular gasoline was also collected from gas stations of the same 3 brands for comparison. GC/MS analysis revealed that biogasoline of 3 different brands was identical, suggesting that adulteration of biogasoline by mixing with regular gasoline was not carried out. ETBE was not detected from any sample of regular gasoline at all. Absence of ETBE in regular gasoline indicates that handling of biogasoline was strictly regulated, making the tracking of biogasoline sample back to its retailer possible.

View full abstract

  Buccal cell samples have become common resources as reference samples in short tandem repeat (STR) typing. In this study, 29 buccal cell samples were collected with 4 kinds of collection kits, and whether the samples provided DNA of sufficient quality and quantity for STR typing was examined. DNA was extracted with EZ-1 DNA Investigator kit from three 3 mm-diameter punches of a Buccal DNA Collector and an EasiCollect, or from the 4 mm tip of an Omni Swab. DNA was eluted from FTA Elute by heat using three 3 mm-diameter punches. DNA concentration was quantified with Real-time PCR and STR typing was performed. DNA concentration ranged from 0.29-2.34 ng/μl for Buccal DNA Collector, 0.57-14.3 ng/μl for Omni Swab, 0.26-3.49 ng/μl for EasiCollect, and 0.04-3.90 ng/μl for FTA Elute. Complete STR profiles were obtained from all the samples of the Buccal DNA Collector, the Omni Swab and the EasiCollect, but only from 25 of the 29 samples of the FTA Elute. Storage at 4°C for 3 days, or 5 times repeated freezing and thawing drastically reduced the concentration of the DNA solution obtained from the FTA Elute. We concluded that the Buccal DNA Collector, Omni Swab and EasiCollect are suitable buccal cell collection kits for STR typing; however, the FTA Elute must be used cautiously.

View full abstract