Syusaku Katsura

Application of ELISA-ABC method to the identification of minute human bloodstains

SummaryBloodstained threads (1 cm in length) were tested to identify human origin by a direct ELISA-ABC method using biotinylated antibody against human HbA0. By this method human bloodstains were clearly distinguishable from bloodstains of other species including Japanese monkey. The minimum detection lirait of bloodstains prepared from undiluted human whole blood was 1 : 5, 120 (28 ng Hb) and that of bloodstains from diluted human whole blood was 1 : 640 ≈ 1 : 1, 280. Human Hb was more easily detectable in bloodstains prepared from diluted human blood after extraction with 5% ammonia than after extraction with phosphate-buffered saline.ZusammenfassungEs wurde eine direkte ELISA-Methode unter Verwendung des Avidin-Biotin-Komplexes zur Identifizierung von menschlichem Blut an einem verdächtigen Blutfleck (ein Faden mit einer Länge von 1 cm) angewandt. In dieser ELISA-ABC-Methode wurde ein Biotin-gebundener IgG-Antikörper (Ziege) gegen Human-HbA0 verwandt. Mit dieser Methode war ein menschlicher Blutfleck von dem Blutfleck japanischer Affen und anderer Tiere sehr leicht unterscheidbar. Die minimalen Grenzen der postiven Reaktionen erwiesen sich beim Blutfleck mit unverdünntem Blut als 1 : 5, 120 (28 ngHb) verdünnte Extraktionsflüssigkeit und beim Blutfleck mit verdünntem Blut als 1 : 640 ≈ 1 : 1, 280 verdünntes Blut. Der Nachweis von Menschen-Hb im Blutfleck mit verdünntem Blut wurde mit 5% Ammoniak-Extraktionsflüssigkeit leichter erbracht als mit Flüssigkeit des Phosphatpuffersalzes.

Rapid and sensitive identification of human blood by an ELISA-ABC method using a biotinylated antibody against human HbA0

SummaryA direct enzyme-linked immunosorbent assay (ELISA) using an avidin-biotin complex (ABC) system for the identification of human blood is described. In this ELISA-ABC method, in which biotin-labeled goat IgG antibody against human HbA0 was used, it was possible clearly to distinguish human blood from the blood of other species, including that of Japanese monkeys. It took about 3 h to obtain the results. Human Hb concentrations ranging from 22 ng to 169 μg produced a positive reaction, and the minimum detection limit in terms of the highest possible dilution of human blood was 1∶640,000.ZusammenfassungBeschreibung einer direkten ELISA-Methode unter Verwendung des Avidin-Biotin-Complexes zur Identifizierung winziger Mengen menschlichen Blutes. In dieser ELISA-ABC-Methode wurde ein Biotin-gebundener IgG-Antikörper (Ziege) gegen Human-HbA0 verwandt. Mit dieser Methode war menschliches Blut von dem Blut japanischer Affen und anderer Tiere sehr leicht unterscheidbar. Positive Reaktionen wurden nach einer Untersuchungszeit von etwa 3 Std. gewonnen. Minimale Hämoglobin-Präparationen zwischen 22 ng und 169 μg erwiesen sich als ausreichend, um eindeutige positive Reaktionen zu erzeugen. Der Test reagierte bis zu einer Blutverdünnung von 1∶640.000-fach positiv.

Human Inter-Alpha-Trypsin Inhibitor (ITI) Silent Allele Found in a Case of Disputed Paternity

Inter-alpha-trypsin inhibitor (ITI) is a 180-kDa glycoprotein having the role of a serine protease inhibitor. It has been revealed that ITI is not a single polypeptide chain structure, but a complex of three kinds of subunits, two heavy (H1 and H2) chains and a light (L) chain (Bourguignon et al. 1983) with chondroitin sulfate cross-links (Jessen et al. 1988). In situ hybridization showed that the H1, H2 and L chains were encoded by separate genes located on chromosomes 3, 10 and 9, respectively (Diarra-Mehrpour et al. 1989). Genetic variation of ITI on an isoelectric point was reported by Vogt and Cleve (1990) being controlled by mainly three alleles (ITI*1, ITI*2 and ITI*3) with autosomal codominant inheritance. The utility of ITI system has been supported from the results of several population studies (Luckenbach et al. 1991; Vogt et al. 1991a, 1991b; Yuasa et al. 1991; Vogt et al. 1992). However, Vogt et al. (1991b) indicated the cautious application of ITI for paternity testing due to an incomplete expression of ITI phenotypes among infants.

Human ZN-Alpha 2-Glycoprotein Phenotyping in Several Populations

Human Zn-alpha2-glycoprotein (ZAG) is widely distributed in various body fluids such as plasma, semen, sweat, saliva, urine and tear, however, the biological function is still unknown. Lately, by means of isoelectric focusing (IEF) followed by immunoblotting, the genetic variation of plasma ZAG has been reported (Kamboh and Ferrell 1986; Nakayashiki and Katsura 1989; Ding et al. 1990). In this study, five new ZAG alleles and the distribution of ZAG alleles in the Japanese, Korean, Philippine, Thai, Brazilian Indian and Papua New Guinean popuations will be reported.