Determination of phenotypes of salivary amylase in liquid saliva and saliva stains

An isoelectric focusing method is described for typing salivary amylase in liquid saliva and saliva stains. The estimated gene frequencies in a British population, calculated on the basis of three alleles operating at a single locus, were Amy 1, 0.909; Amy 2, 0.065; Amy 3, 0.026. This system may be useful in forensic investigations.     

Determination of MN blood group from blood stains by electrophoresis and immunoblotting

The determination of blood groups from blood stains is extremely important in medicolegal practice, but there is the possibility of an error in the determination of MN phenotypes by the absorption-elution test. We investigated a new method applying electrophoresis and immunoblotting. As a consequence of various experiments, the most appropriate pretreatment of blood stains was as follows. Blood stains were immersed in physiological saline for 0.5 to 1 h and centrifuged. The supernatant was discarded. The sediment was dissolved in sample buffer (TRIS-buffered physiological saline containing 2% sodium dodecyl sulfate) and followed by thermodegradation. It was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After transfer to a nitrocellulose membrane by Western blotting, MN phenotypes could be determined accurately from blood stains by an enzyme immunoassay (EIA) using commercially available polyclonal anti-M and anti-N sera. For blood stains more than 1 month old it was not easy to determine the MN phenotypes.     

Detection of amphetamine in bloodstains,semen, seminal stains,saliva, and saliva stains

Low nanogram quantities of amphetamine were detected in 100μl samples of dried bloodstains using radioimmunoassay. Saliva, saliva stains, semen, and seminal stains also contained measurable quantities of the drug.     

Detection of Lea substance in saliva stains by enzyme-linked immunosorbent assay (ELISA) using anti-gum arabic serum

It is known that rabbit anti-gum arabic (GA) serum has cross-reactivity with Lea antigen, and that, by using this cross-reactive anti-Lea antibody, the presence of Lea antigen in red blood cells and saliva can be demonstrated with accuracy. We have devised a rapid and highly sensitive method for detecting Lea substance in human saliva by the enzyme-linked immunosorbent assay (ELISA) method using an anti-Lea antibody isolated from anti-GA serum by affinity chromatography on Synsorb Lea. The ELISA plate, coated with the specific anti-Lea antibody, adsorbed the Lea substance in saliva which was subsequently identified by adding enzyme labeled anti-Lea IgG in that order. The method could detect the Lea substance in Le(a+) saliva stains as small as 0.1 by 0.1 cm in size that had been stored at room temperature for three weeks and in Le(a+) saliva stains 0.7 by 0.7 cm in size that had been stored for ten years. This method seems to be useful for quantitative analyses of the Lea substance in various body fluids.     

Lewis typing of human saliva stains by enzyme-linked immunosorbent assay (ELISA) using monoclonal anti-Le(a) and anti-Le(b) antibodies

The Lewis blood grouping of human saliva stains could be detected by an enzyme-linked immunosorbent assay (ELISA) using anti-Le(a) and anti-Le(b) monoclonal antibodies with an avidin-biotin complex (ABC). The saliva stains (1.0 by 1.0 cm in size) were used as samples and not only could the Lewis substances of 57 individual stains be correctly typed by this method, but also it was clarified that there are several different secretion patterns of amounts of Le(a) and Le(b) substances in 3 individual Lewis types.     

Identification of saliva stains by determination of the specific activity of amylase.

The specific activity (enzyme activity/protein concentration) of amylase was determined for the identification of saliva stains. The specific activity of amylase in saliva stains rapidly decreased during the first hour but, from 1 to 28 days, this decrease was much less when the stains were kept at room temperature. Stains of various human biological materials, breast milk, nasal secretion, meconium and vaginal secretion showed comparatively high amylase activity, but the saliva stains could be differentiated by their high specific activity of amylase, over 2 I.U./mg. When saliva stains were contaminated with blood or vaginal secretions at various ratios, the specific activity of amylase decreased with increase in the ratio of contaminant, especially when the contaminant was blood. However, the specific activity of amylase was still higher than 2 I.U./mg even after one fifth volume of blood was added or after five volumes of the extract of the stains of vaginal secretions were added.     

Determination of phenotypes of haptoglobin and GLM(1) factor of human GM system in stains with admixtures of blood of some fish strains

It was shown that the phenotypes of haptoglobin (Hg) can not be detected in stains of dried blood of salmon, roach and bream. The results of experimental research of blood of the above fish species are described according to the Cm system. It was proven as possible to identify the human blood in stains with admixtures of blood of the above fish by using the Hp and Cm systems.     

Detection of orosomucoid 1 phenotypes in semen and semen stains

Orosomucoid 1 phenotypes were detected in seminal plasma by isoelectric focusing and immunoprinting. The orosomucoid 1 phenotypes in seminal plasma correlated with the types found in the corresponding serum specimens. Semen stains stored for ten days could be typed for orosomucoid 1. The present work revealed that orosomucoid 1 is a useful genetic marker for the medicolegal grouping of semen stains.     

The use of monoclonal antibodies in analyzing blood and saliva stains by the mixed agglutination reaction

Possible use of monoclonal antibodies anti-A, anti-B, anti-M and anti-N in mixed agglutination reaction during the analysis of blood and secretion traces of different age is demonstrated. Dependence of antigen A and B detection in salivary traces on secretory type was determined.     

Further evidence for phenotypes and gene frequencies of nine salivary polymorphisms in Japanese population

Nine salivary polymorphic systems (Pa, Pb, Pr, Db, PmF, PIF, Ph, Amy1 and s-AcP) were examined using parotid and whole saliva from random Japanese individuals. The gene frequencies obtained were: Pa+ = 0.221, Pb1 = 1.000 Pr1 = 0.741, Db+ = 0.033, PIF+ = 0.715, Ph+ = 0.029, Amyv1 = 0.013 and s-AcPA = 0.217, respectively.     

The establishment of the presence of feces in stains by agar gel electrophoresis]

An electrophoretic method of feces determination in stains was developed. Method is based on detection of acid phosphatase only in feces since it is not detectable in human blood and other secretions by this method.     

Allele distribution of 15 PCR-based loci in the Rwanda Tutsi population by multiplex amplification and capillary electrophoresis

A genetic study of 15 autosomal STRs is carried out (D2S1338, D3S1358, D5S818, D7S820, D8S1 79, D13S317, D16S359, D18S51, D19S433, D21S11, CSF1PO, FGA, TPOX, THO1, VWA) in a sample of unrelated Tutsis. The molecular phenotypes were determined by means of multiplex strategies (AmpFlSTR Identifiler PCR Amplification Kit, Applied Biosystems) followed by capillary electrophoresis.     

Studies on glyoxalase 1 isoenzymes in semen stains: polymorphism in Himachal (India) population

One hundred and ten pairs of blood and semen samples and their stains were studied to type glyoxalase 1 (GLO 1) isoenzymes using agarose-starch medium. A good agreement was observed between the phenotypes expressed in blood and semen samples of the same donor. No GLO 1 activity however could be demonstrated in the vaginal swabs tested. The gene frequencies of GLO 1 polymorphs in Himachal population has been worked out and their stability studies carried out at -12 degrees C and at room temperature.     

Detection of hemagglutinins in dried saliva stains and their potential use in blood typing

Since 1928, hemagglutinins have been known to exist in saliva; however, they have not been utilized as evidence in criminal investigations because in the past, techniques for measuring them have not been sufficiently sensitive. In this paper we describe improved techniques for detecting salivary hemagglutinins and report initial results obtained with these methods. The stability of salivary hemagglutinins at several different temperatures was examined in liquid samples and in dried stains on filter paper, cigarette butts, and envelope flaps. Our observations indicate that salivary hemagglutinins may be sufficiently stable, over periods of one to several days at ambient room temperatures, to be of value to forensic science investigators. The results of the hemagglutinin assay are not affected by the age or sex of the sample donor. Because salivary hemagglutinins can be used to determine ABO blood type, analyses of this kind can serve as an important confirmatory test which the forensic serologist can use in conjunction with salivary agglutinogen determinations.