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PubMed1982

[The use of remission analysis for direct colorimetric determination of age of blood stains (author's transl)].

A review is given of the conventional methods of determining the age of blood stains by forensic examination of trace evidence. Detailed analysis of the samples in several steps, which destroyed the traces, and the widely differing results do not appear satisfactory with regard to their use in forensic investigations. Considering the change in color with increasing age the three coloric parameters (chroma, hue, and value of the blood stain) can yield exact numerical measurements which allow to draw relatively reliable conclusions as to the age of the blood stain. Of the three colorimetric methods, based on equalization, three regions, and spectral analysis, respectively, only the last may be considered really satisfactory and up to the requirements of forensic medicine. It has the additional advantage of being independent of the visual acuity of the examiner and of the type and quality of the source of illumination. The aging of freshly taken venous blood, which has been deposited on filter paper or linen, can last up to 33 days, either in the dark or in day light, with due regard to temperature and humidity. The spectrophotometric color measurements taken place on the immediate surface of the blood stain in the range of the visual spectrum between 400 and 600 nm. The continuously drawn spectral curves are then subject to remission analysis the first step. The second step consists in a mathematical computer analysis of color valence measurements.

Blood Stains↗
PubMed1999

Detection of epithelial cells in dried blood stains by reverse transcriptase-polymerase chain reaction.

The identification of menstrual blood stains can be improved by detection of messenger ribonucleic acid (mRNA) specific for epithelial (endometrial) cells. RNA molecules, however, are believed to be unstable and require careful sample processing. In this study, we have investigated the extraction of RNA suitable for reverse transcriptase-polymerase chain reaction (RT-PCR) from dried blood stains stored for up to six months. With a modified RNA isolation protocol, it was possible to obtain RNA from dried blood stains with at least 5 x 10(2) leukocytes. In an additional experiment, we evaluated the RNA isolation from mixed stains composed of leukocytes and T47D cells, a breast cancer-derived cell line with epithelial origin. Detection of 10(2) T47D cells in a total number of 10(5) leukocytes was possible by amplification of cytokeratin 19 mRNA and progesterone receptor-mRNA specific for hormonally regulated epithelial cells. In both experiments amplification results were not dependent on storage time with similar data from one day to six months. Furthermore, it was possible to identify dried menstrual blood samples by showing the presence of mRNA specific for epithelial cells. These results demonstrate for the first time, that RNA suitable for RT-PCR, can be isolated from forensic specimens stored up to at least six months, and that a small number of epithelial (endometrial) cells can be identified in dried blood specimens. Using this method, it will be possible to identify the origin of small and partially degraded blood samples which can be especially useful in forensic evaluation of cases with sexual offense.

Base Sequence↗
PubMedDate not supplied

Streptokinase versus alteplase and other treatments for acute and delayed thrombolysis of blood stains in clothing.

OBJECTIVE: To assess the usefulness of heparin, alteplase, and streptokinase in removing blood stains. DESIGN: Randomised controlled trial. SETTING: Hospital laundry. INTERVENTIONS: Blood stains were allocated to treatment with alteplase, streptokinase, heparin, a commercial enzymatic stain remover, or no treatment at all after three or seven hours and then washed in hot or cold water two hours later. RESULTS: Both hot water and early treatment were strongly associated with improved stain removal. All four treatments were associated with a worse outcome than no treatment at all, although for streptokinase this trend did not reach significance. The commercial stain remover gave the worst results of all treatments tested. CONCLUSIONS: Contrary to popular wisdom, hot water is much more effective than cold in removing blood stains. Methodologically rigorous research and evidence based principles are needed within the laundry industry, and the role of thrombolytic drugs should be assessed further.

Blood Stains↗
PubMed1977

[A photometric method for the diagnosis of pregnancy in blood stains by determination of heat-stable alkaline phosphatase (EC 3.1.3.1). A modification of Oya, Asano and Fuwa's technique (author's transl)].

A quantitative method, described by Oya and coworkers was modified. In this way, venous blood from pregnant and nonpregnant women in blood stains can be reliably differentiated up to at least 19 months storage although as a qualitative test only. 1--3 cm2 of cloth stained with blood is needed and even less from placental blood stains.

Alkalies↗
PubMed1992

Detection of human proteins in buried blood using ELISA and monoclonal antibodies: towards the reliable species indentification of blood stains on buried material.

The survival of human proteins in blood stains on fragments of cloth buried in exposed soil was examined in a 15-month investigation carried out from September 1990 to December 1991. During this period there was a wide variety of weather conditions. Samples were exhumed at 4-weekly intervals for 16 weeks and finally at 65 weeks; extracts of the stains were tested for albumin and IgG using a highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) performed with monoclonal antibodies. Human albumin survived well throughout the 15 months of study, but IgG could be detected only in the 4- and 8-week samples. The reactions for IgG were weaker than those for albumin, although the method's sensitivity (10 ng) was the same for each protein. Appropriate buried and non-buried control experiments were carried out using cloth, either unstained or stained with human blood or animal sera; there was no cross-reactivity between human and the other species investigated and soil did not affect the assay; under laboratory conditions, IgG and albumin survived equally well. The system's versatility was illustrated by using monoclonal anti-bovine-albumin to detect specific albumin in the extracts of buried cloth which has been stained with bovine serum. It was concluded that ELISA performed with monoclonal antibodies could be of great value in identifying blood stains for forensic purposes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Study on the factors affecting ABO grouping of blood stains.

The Biologist in forensic work is confronted with many problems on grouping of dry blood stains, these problems are due to many factors and environmental changes which may mislead the analysis. The present work deals with the factors affecting ABO grouping of dry blood stains in Riyad, including exposure to extremes of temperature, from refrigeration at -4 degrees C up to heating at 150 degrees C, effect of time till 6 months, occurrence of the stains on different fabrics, and effect of putrefaction. The results showed that, ABO grouping of dry blood stains was not affected by different substrata or cloth fabrics. 30% of the samples gave erroneous results when exposed to temperature at 150 degrees C for 2 hours. Storage for 6 months at room temperature ranged from 29 degrees -44 degrees C, representing the ambient temperature met with in Riyad, gave no effects. Putrefaction showed that the antigens were deteriorated in 80% of cases after 90 days. Chromatographic separation was a suitable procedure before grouping the samples which were buried in sand or mud.

ABO Blood-Group System↗
PubMed2000

Errors in ABO typing of blood stains using PCR.

Genotypes of the ABO blood group system were investigated using a multiplex PCR and subsequent restriction enzyme digestion on experimental blood stains. Differences were found when typing blood between the PCR and serological methods and one blood sample, typed as B with the agglutination test was classified as AB using the method described here. The subsequent sequencing procedure revealed the genotype to be BB. Methodological causes for errors in typing which should be taken into consideration are discussed.

ABO Blood-Group System↗
PubMed1994

A novel assay for typing Rh antigens in blood-stains using a lectin specific to the bisecting N-acetyl-D-glucosamine side chain of glycoprotein.

A unique sandwich enzyme-linked immunosorbent assay for the determination of Rh antigens in blood stains has been developed using Rh antisera and phaseolus vulgaris E4/peroxidase conjugate (PHAE4/PO). The appropriate antiserum for detecting Rh C, c, D, E or e was coated on the inner surface of microplate wells, and the sample antigens from blood stains, solubilized with n-octyl-beta-D-glucopyranoside, were then placed in the wells. After washing the wells repeatedly, PHAE4/PO was added. Bound PHAE4/PO was detected by the development of colors using o-phenylenediamine/H2O2. All Rh antigens corresponding to the antisera were clearly detected using this technique. The detection limit expressed by sample dilution was more than 2 x 10(5) times (volume/dried blood weight) for the various antigens from the fresh 5 x 5 mm2 blood stain. Even when the blood stain samples were left beside a sunny window at room temperature for 2 months, Rh antigens were still detected. When the ABH, MN, P1, Kidd, Duffy and Lewis blood grouping systems were tested with similar ELISA procedures PHAE4 did not recognize any antigen. Since PHAE4 specifically recognizes and combines with the bisecting N-acetyl-D-glucosamine side chain, it was concluded that the glycoprotein was a component of all Rh antigens immune complexes.

Acetylglucosamine↗
PubMed1985

Corneal blood staining at low pressures.

After sustaining blunt ocular trauma and total hyphemas, two patients quickly developed corneal blood staining in the absence of elevated intraocular pressure. Corneal blood staining has previously been reported to occur in association with hyphema and raised intraocular pressures. Although several bibliographic references acknowledge that corneal blood staining may occur without raised pressure, a careful review of the literature failed to reveal any well-documented cases.

Adult↗
PubMedDate not supplied

[Immunoelectrophoresis used for identification of blood stains in forensic medicine].

Conclusive evidence was obtained in this study that immunoelectrophoresis could be used in the identification of blood-stains in two particular cases: for diagnosis or approach to diagnosis of the age of a blood-stain, and for diagnosis of the human origin of a blood-stain that had been treated with petroleum products. Although many aspects should be investigated in more detail, it is certain that this method can be employed in combination with other methods.

Blood Stains↗
PubMed1981

Microspectrophotometric studies of Romanowsky stained blood cells. II. Comparison of the performance of two standardized stains.

This paper describes a comparison of the performance of two standardized Romanowsky blood stains, namely those of Marshall et al. and Wittekind et al., both containing azure B and eosin alone. Stain performance is assessed objectively by the use of three complementary techniques, all based on the visible absorbance spectra of stained cellular substrates. The first of these techniques is a simple comparison of the shapes and heights of the absorbance spectra. The second technique uses the CIE Colorimetric System, and thus permits the quantitation of colour in a manner that agrees with human observation. CIE co-ordinates (chromaticity points, luminance) are calculated directly from absorbance spectra. The third technique is that of spectral subtraction, which yields a set of factors which describe the quantities of component dyes which are bound by the object. This technique, unlike the other two, requires a priori knowledge of the dyes used in the stains, and their spectra when bound to cellular substrates. Although the differences between the two methods are subtle, and hard for the subjective observer to define, the objective methods described here do show statistically significant differences. Wittekind's stain produces less intense staining, except for lymphocyte and monocyte cytoplasms. To the human eye, the differential coloration of these two substrates is more pronounced, but the difference between all nuclei and cytoplasm is less marked. The major difference in the uptake of dye components is in the small quantities of eosin dimer that are bound in this technique.

Blood Cells↗
PubMed1987

Detection of AB antigen in blood stain using enzyme-linked immunosorbent assay.

A new method for detection of AB antigen from blood stain using enzyme-linked immunosorbent assay (ELISA) is described. Flat-bottomed-wells of polystylen plate coated with human anti-B or rabbit anti-A were sensitized with AB antigen which was extracted from blood stain with 1% octyl-glucopyranoside in 0.1 M phosphate buffer pH 8.0. Mouse monoclonal anti-A or anti-B, and peroxidase conjugated anti-mouse immunoglobulin were added to the wells, respectively. Subsequently, the substrate was dropped into the wells, and the absorbance of the solution was measured. By this method, we could distinguish AB group blood stain from the mixed stain of A and B group bloods. When rabbit antiserum was used as the first antibody, differentiation between these antigens was unsuccessful presumably because of non-specific adsorption.

ABO Blood-Group System↗
PubMedDate not supplied

[The use of proteases for enhancing the sensitivity of reactions to detect group isohemagglutinins in the forensic medical study of blood stains].

Group a and b isohemagglutinins are much better detected in blood stains by enzyme-treated, but not native red cells. This is specially true for antibodies of poorly dissolved strains, whose group antibodies can be detected virtually only by enzyme-treated red cells in papain extracts possessing the highest immunological activity. Two sensitive methods for the detection of isohemagglutinins have been developed: extract titration and modified Lattes' method, permitting a reliable detection of group antibodies in old blood stains, undetectable by routine methods.

ABO Blood-Group System↗