Determination of ABO blood group markers in single hair segments

Determination of ABO-blood group on human hair segments of 79 individuals was performed by means of modified Yada absorption-elutions technique. The studies were carried out with 4 micron thin cross sections of hair. In 79% of the cases it was possible to establish the correct blood group. Individual hair segments of 0.5 cm length yielded reliable results. Antigen identification was best with blood group B and most difficult of the AB type.     

Immunochemical studies on blood group A substance from human hair

Blood group A-active substance was extracted from urea-treated human hair uith methanol-ethyl ether 1:1, v/v) or chloroform-methanol (1:1, v/v). The serological activity of blood group A substance in the hair was destroyed by A-decomposing enzyme from Clostridium tertium with concomitant development of blood group H activity. It is concluded therefore that the extract from the hair of group A contained blood group A-active glycolipid with N-acetylgalactosamine as the non-reducing sugar.     

The ABO blood grouping of a minute hair sample by the immunohistochemical technique

The unlabeled antibody (PAP) immunoperoxidase technique was applied to the ABO blood grouping of human scalp hairs. Hair samples were subjected to longitudinal- or cross-sectioning, thus obtaining suitable samples for subsequent immunostaining. The immunostaining was carried out using rabbit anti-A and anti-B sera as the primary antibodies. With this technique, the group-specific staining which is revealed as a dark brown precipitate was clearly observed within the medullae of the hair shaft, and depending on the presence or absence of these precipitates, respective blood groups of unknown hair samples were determined. At the hair root, on the other hand, positive stainings were observed not only in medullary cells but also in some cortical cells of the keratogenous zone. From the present study, it can be safely said that this technique is of practical use for the ABO blood grouping from a minute (less than 3 mm) hair sample.     

Determination of ABO(H) blood group substances from finger and toe nails

Thirty-six finger and toe nails were analyzed for ABO(H) blood group substances by the modified absorption elution method. The blood groups from nails were successfully determined in all the samples.     

Examinations of ABO bloodgroupings of human hair

In order to bring into play the function of human hair ABO bloodgrouping in the field of medicolegal expertise on material evidence and raise its accuracy, the author, through a test on human hair of 500, makes some emphatic researches on the disposal of hairs, on the application of anti-A and anti-B serum, on the selection of red blood cell indicator and on the elution temperature as well. Five hundred samples of human hair have been examined upon the basis of the improved operation method and through the application of anti-A and anti-B serum, the titer of which being 1:128, and therein, fine results have been obtained.     

Effect of blood group active micro-organisms on the ABO grouping of human whole saliva

Three saliva samples with false positive ABO grouping results were assayed for blood group active organisms, using a variety of selective media to isolate representative strains from the salivary microflora. Eight out of 40, 8 out of 40 and 4 out of 30 strains from the three samples, respectively, showed blood group activity, which correlated well with the false positive specificities of the saliva samples. In all cases the false reaction only lasted a few days. Investigation of one of these samples before and after the appearance of the false positive activity yielded only one out of 40 blood group active organisms, using the same methods. Similar investigation of two "normal" saliva samples found none out of 40 and one out of 40 blood group active organisms, respectively. It is concluded that occasional false positive ABO grouping reactions of saliva samples are probably caused by the presence of unusually high numbers of blood group active micro-organisms, due to disturbances in the ecological balance of the salivary microflora.     

Immunohistochemical study of the pattern of distribution of ABO blood group substances in male genitalia

The distribution of the ABH antigens was investigated in 11 different sections of the male genitalia in 53 autopsy cases; the peroxidase-antiperoxidase technique was used. Specific staining of the epithelia differed considerably among and between individuals. Nonsecretors showed a tendency toward less staining in the epithelia, whereas in the endothelia there was no difference. A2 cases could be recognized, as the endothelia were marked almost completely with anti-H. In A2B nonsecretor epithelia and endothelia, there was only a minimal reaction with anti-A and anti-H. Spermatozoa were irregularly stained in the ampulla of the vas deferens, whereas in the testis and epididymis no reaction could be found.     

Immunoenzyme AB0 blood group determination using a single hair. I

The immunoenzyme technique was used to determine the ABO blood group of strands of human scalp hair. The hair was obtained from 168 individuals of known blood groups (A1: n = 58; A2: n = 11; B: n = 28; O: n = 46; A1B: n = 16; A2B: n = 9). Immunostaining was carried out by using monoclonal anti-A, anti-B and anti-H as primary antibodies. Group-specific staining was clearly observed within the medulla of the hair. The ABO blood group of all hair samples was determined correctly by the Sternberger (PAP) or APAAP (immunoalkaline phosphatase) technique. The present study indicates that immunoenzyme techniques can be regarded as practical methods for determining ABO blood group of hair.     

Interlaboratory comparison of quantitative determinations of drug in hair samples

Testing human hair for drugs of abuse is a relatively new technique which requires control before being fully accepted in Justice applications. A consensus procedure was recently proposed to the four French Laboratories performing hair analysis for opiates and cocaine. Results of two independent controls have shown that the laboratories have performed very well quantitatively, using the recommended method. In order to compare these results with those obtained by other procedures, one sample was sent to 15 laboratories concerned with the analysis of human hair for drugs of abuse in Germany, Italy, Spain, and United States. Results from this study have indicated that the French recommended method is in accordance with the general procedures.     

Determination of hair group by preserved hair cell bulbs

Difficulties appearing in determining hair group were the reason for conducting this research for the purpose of finding additional possibilities to solve the mentioned problem. It was established that the preliminary cell coloring, related with determination of hair sex, does not influence a consequent detection of antigens. Methods or the fixation of material were selected. The most suitable reagents and their titers as well as different time periods of absorption for detecting antigens A and B are offered. All stages of examination are described in detail.     

Heteroplasmy in hair: differences among hair and blood from the same individuals are still a matter of debate

The analysis of mitochondrial DNA (mtDNA) is a useful tool in forensic cases when sample contents too little or degraded nuclear DNA to genotype by autosomal short tandem repeat (STR) loci, but it is especially useful when the only forensic evidence is a hair shaft. Several authors have related differences in mtDNA from different tissues within the same individual, with high frequency of heteroplasmic variants in hair, as also in some other tissues. Is still a matter of debate how the differences influence the interpretation forensic protocols. One difference between two samples supposed to be originated from the same individual are related to an inconclusive result, but depending on the tissue and the position of the difference it should have a different interpretation, based on mutation-rate heterogeneity of mtDNA. In order to investigate it differences in the mtDNA control region from hair shafts and blood in our population, sequences from the hypervariable regions 1 and 2 (HV1 and HV2) from 100 Brazilian unrelated individuals were compared. The frequency of point heteroplasmy observed in hair was 10.5% by sequencing. Our study confirms the results related by other authors that concluded that small differences within tissues should be interpreted with caution especially when analyzing hair samples.     

Rapid direct PCR for ABO blood typing

Many different molecular typing methods have been reported to complement routine serological ABO blood typing in forensics. However, these ABO genotyping methods are often time-consuming and call for an initial DNA isolation step that requires the use of expensive kits or reagents. We report here a rapid direct ABO genotyping method that eliminates the need for DNA extraction from fresh blood, hair, and body fluid stains before PCR. Using a fast PCR instrument and an optimized polymerase, the genotyping method-which employs a multiplex allele-specific primer set for the simultaneous detection of three single-nucleotide polymorphism (SNP) sites (nucleotides 261, 526, and 803)-identifies A, B, O01/O02, O03, and cis-AB01 alleles in around 70 min from sample collection to electropherogram. Not only will this ABO genotyping method be efficiently used in forensic practice for rapid screening of samples before full-blown multilocus short tandem repeat profiling, but it will also demonstrate an example of rapid direct genotyping of SNPs that offers the advantages of time- and cost-efficiency, convenience, and reduced contamination during DNA analysis.     

Dependability of ABO findings in stored blood samples

Report on the occurrence of "acquired" A or B in stored blood samples. This bacterial alteration is of importance when an indirect experimental investigation with the absorption-elution technique is needed in advanced cases of hemolysis. One has to consider this disturbing factor in identification tests (alcoholic blood samples). In dried blood stains we did not notice this problem, but it has to be taken into account in genitals stains.