Identification of characteristics in blood and semen stains--a review

A review is given of proved techniques for detection of individual characteristics in stains of blood and semen. Above all the results of German investigators are considered because these have mostly been insufficiently treated in the English-language literature, for instance papers concerning Gm and Km typing.     

Gm(1,2,4,10,21) and Km(1) factors in vitreous humor

The presence of Gm(1,2,4,10,21) and Km(1) factors in vitreous humor taken from human corpses was investigated. The results revealed a good agreement between the factors detected in this biological material and in blood. Their presence in vitreous humor is independent of the secretor type.     

Gm typing by enzyme-linked immunosorbent assay (ELISA)

A solid-phase ELISA for Gm typing is described. A mixture of anti-Gm serum (or monoclonal anti-Gm antibody) and test serum was incubated in microtiter wells coated with IgG or its fragments of appropriate Gm type. After washing of the wells, the bound antibody was detected with peroxidase-labeled second antibody. The Glm(3), G3m(16), and G3m(21) antigens could be identified by this technique. Since some of the human anti-Gm sera and anti-Rh0 sera required for the conventional hemagglutination-inhibition method are hard to obtain, the ELISA system using anti-Gm antibodies and no anti-Rh0 sera may serve as an alternative to the conventional method.     

Genetic markers in human bone: II. Studies on ABO (and IGH) grouping.

A combination absorption-elution, two-dimensional absorption-inhibition procedure was used to determine the ABH antigen composition of a series of human bone specimens of known ABO type that had been aged up to nine months under dry and humid conditions at ambient temperature, 37 degrees C, and 56 degrees C; at ambient temperature in dry and wet soil; and buried in soil outdoors. Grouping data for the separate elution and inhibition testing, as well as for the combination procedure, are given. The combination method was found to be a highly reliable procedure for bone tissue ABH typing. Some data on microbial contaminants of human bone specimens aging in soil, and their effects on ABH typing results, are presented. No direct correlation between the properties of microbial contaminants and specific changes in the ABH antigenic composition of aging bone tissue specimens could be ascertained. Data on IGH antigen determination and on the quantitation of immunoglobulin G (IgG) in human bone tissue extracts indicated that immunoglobulin levels were typically too low to expect routinely successful Gm antigen testing results. However, these factors can sometimes be determined in fresh bone tissue extracts, particularly if the extracts are concentrated.     

血痕Gm(2)因子检验及分布

应用抗 Gm(2)血清和抗 D-Gm(2)血清以及 Gm 标准对照血清,选择新鲜 ORh(D)红细胞,用凝集抑制试验,对北京地区269人血清或新鲜血痕进行检验。结果 Gm(2+)分布为29%,Gm(2-)分布为71%。对已确定 Gm(2)型的230份血痕,在室温放置14个月后再次检验,结果与第一次所测血型完全相同。同时对11起案件中的血痕检验了 Gm(2)因子,起到了排除嫌疑人或增加与案件相关联的作用。     

Gm(11) grouping of dried bloodstains

An absorption inhibition method for the detection of gamma marker Gm(11) in dried bloodstains is described. Particular reference is made to the association of Gm(11) with Gm(-1, -2). When a dried bloodstain fails to inhibit anti-Gm(1) and anti-Gm(2), this may represent a true Gm(-1, -2) result or there may be insufficient material to inhibit either antibody. The detection of Gm(11) in a bloodstain extract provides an objective means of confirming the apparent absence of Gm(1) and Gm(2) as representing a true Gm(-1, -2) result. This antigen compares very well with other blood group systems with regard to the amount of bloodstain required for analysis and its stability. No evidence is available for preferential loss of Gm(1) and Gm(2) relative to Gm(11) in dried bloodstains.     

Gm and Km frequencies in West Germans: usefulness of typing Gm(1,2,3,10,21) and Km(1,3) in paternity cases

The immunoglobulin allotypes G1m(1,2,3) and G3m(10,21) were typed in 2855 unrelated West German adult individuals. 1455 individuals were typed for the factors Km(1,3). Phenotype and haplotype frequencies are reported. The usefulness of this routine typing programme in paternity tests is demonstrated in three case reports.     

Possible common partial antigens in human Ig allotype structure and ubiquitous bacteria, studied with the example of Escherichia coli

As can be learned from the literature, bovine serum may contain antibodies directed against human immunoglobulin allotypes. This gave rise to the question of what the origin of those antibodies is. We tested bacteria (E. coli) by means of the haemagglutination inhibition assay, which is used to type either Gm or Km factors. Anti-G1m(2) and anti-G3m(10)-specific antibodies were inhibited by the bacteria in a clear-cut manner, as was anti-Km(1), albeit less significantly. In contrast, the bacteria tested almost totally failed to inhibit anti-G3m(21) serum. The results lead to the assumption that E. coli may carry both Gm- and Km-like antigenic structures, which are presumably the antigenic material leading to immunization of cattle. Furthermore, new attention is drawn to a mechanism for immunization which is discussed regarding the genesis of either AB0 isoagglutinins in man or other "naturally occurring" antibodies.     

Sensitivity of various study technics in blood stains

Quadratic pieces of fleece measuring 16 mm2 were soaked with 10 different blood-samples in the dilution steps of 1:1, 1:10, 1:100, 1:1000, respectively, and were tested in blood group typing and identification tests of forensic serology. The above spezified dilutions correspond with 5 microliters, 0.5 microliter, 0.05 microliter and 0.005 microliter of blood, respectively. The detection limit of the microspectrometric test for blood was the dilution 1:10, of the porphyrine test a dilution above 1:100, whereas the preliminary test for blood (peroxidase) succeeded always up to a dilution of 1:1000 and the species determination by the radial immunodiffusion test in agar gels succeeded in most cases op to a dilution of 1:1000. The detection limit of the anti-human globulin inhibition test was between the dilution steps 1:10 and 1:100 when non-titrated and undiluted anti-human globulin serum was used. Gc- and ABO-grouping were possible up to a dilution of 1:100 and were thus the most sensitive grouping systems. Phenotyping of the enzyme-systems and the Gm/Km-system usually required stains with considerably higher blood concentrations i.e. stains of undiluted blood.     

Trial elution of semen stains and measurement of IgG level: a contribution to Gm typing

The conditions for the elution of IgG in seminal stains have been investigated systematically. The amount of IgG recovered could neither or hardly be influenced by variation of the time (15 minutes to 120 hours) and temperature (4 degrees C, 20 degrees C, 37 degrees C, 56 degrees C) of elution, nor by mechanical treatment (cutting in small pieces, crushing), ultrasonic treatment or addition of a detergent. For fresh traces and such of an age of several weeks an elution time of 30 minutes at room temperature is sufficient; for very old stains an elution up to 2 hours may be recommended for safety. The investigations were restricted to the measurement of the IgG concentration in the eluates. No statement can yet be given about the biological value of the IgG for Gm typing due to this investigation alone.     

Blood grouping in a sexual assault case: criteria and methodolgy for genetic marker analysis

A sexual assault case was received in the laboratory. Upon examination, a small bloodstain was located on a bed sheet that was recovered from the defendant's motel room. Typing the whole blood samples from the defendant and the victim revealed that both blood samples exhibited identical phenotypes in eleven different genetic markers. Gm(1) and Gm(2) analysis was then performed on the two whole blood samples which provided discrimination between the two parties.     

不同分型方法的STR分型差异

目的调查不同的STR分型系统之间分型的一致性。方法 10 0例不同个体的DNA样本分别用单位点聚丙烯酰胺凝胶银染法和PowerPlex16System试剂盒对 13个法医学常用STR位点进行基因分型 ,并比较两种不同分型系统间的分型结果。结果 1例样本在D8S1179位点出现了分型不一致的结果 :银染法的基因型为 12 / 14 ,而用PowerPlex16System试剂盒的分型则为 12 / 15。结论不同的STR分型系统可导致不同的基因分型     

A new 39-plex analysis method for SNPs including 15 blood group loci

A novel 39-plex typing system for single nucleotide polymorphisms (SNPs) has been developed. This multiplex approach has the advantage of being able to type 38 autosomal SNPs and one sex-discriminating base exchange site on the X and Y chromosomes rapidly and simultaneously. The SNP loci on the autosomes, which we examined, contain 15 loci distributed on blood type genes: three on RhCE, two each on Km and Gc, and one each on Duffy, AcP1, Tf, MN, GPT, EsD, PI, and Kidd genes. Thirty-seven genomic DNA fragments containing a total of 38 SNPs and one sex-discriminating site were amplified in one multiplex PCR reaction. Following the reaction, single nucleotide primer extension reaction was performed by dividing these SNP loci into five groups. The SNP type of each of the 39 loci was determined at one time by capillary electrophoresis using the newly designed multi-injection method. The combined PD (power of discrimination) of this typing system was (1-1.1) x 10(-14), and the MEC (mean exclusion chance) was 0.9990. We applied this system to forensic cases, including 16 paternity testing cases (13 non-exclusion and three exclusion cases) and one personal identification case. For the paternity testing cases, the highest Essen-M?ller's W-value was 0.9999995. The pM (matching probability) of the personal identification case was 2.22 x 10(-17). These data showed that this system was an excellent tool for use in forensic cases of paternity testing and personal identification.     

Incidence of GM-system factors in indigenous peoples of Buryatia

Distribution of blood groups by the serum Gm system was studied in Buryats and in the entire population of Buryatia without consideration for the national appurtenance and with consideration for sex. Differences in the incidence of the factors were detected.     

Km(3) identification by enzyme-linked immunosorbent assay (ELISA) as an internal control for Km(1) activity determined by inhibition in dried bloodstains

A stability study comparing the identification of kappa marker Km(1), using the classical inhibition of agglutination, and the identification of Km(3), using an automated enzyme-linked immunosorbent assay (ELISA) technique, was done. Preliminary tests were performed to establish the specificity and sensitivity of the methods. Based on the results, the quantities of stain required to detect each marker were determined. Blood samples from 24 staff donors of known phenotype were aged at room temperature and at 37 degrees C in the dried stain and liquid forms. In addition, 192 stains from cases 1 to 7 months old and 76 staff-donor stains from 1 1/2 to 10 years old were tested in dried stain form. The known sensitivity of the ELISA technique was exploited by deliverately testing a decreased quantity of antigen. As control stains were aged beyond the detectable limits of sensitivity, results consistently showed an almost simultaneous success or failure to detect Km(1) and Km(3). This indicates that the interpretive criteria established for ELISA are sufficiently demanding to eliminate the danger of reporting false Km(-1) results but true Km(3) results.     

Effect of the genetic markers Kell(K1), P1, Km(1), Tf(C

E Osterhaus  P Birkner 《Zeitschrift für Rechtsmedizin》1984,91(3):231-234

Development of a 24-plex Y chromosomal STR loci typing system

Y-chromosomal short tandem repeat (Y-STR) loci can play important roles in forensic casework and paternity testing. In our paper, 24-plex Y-STR typing system, which includes 3 loci (DYS635 and DYS385a/b) existed in current widely available commercial kits and 21 additional loci (DYS531, DYS630, DYS622, DYS552, DYS510, DYS449, DYS459a/b, DYS446, DYS443, DYS587, DYS527a/b, DYS460, Y-GATA-A10, DYS520, DYS557, DYS522, DYS481, DYS570, DYS444) was established with 5-dye fluorescence labeling. 200 unrelated Chinese Han males were successfully genotyped with the system and 198 haplotypes were observed. The gene diversity of each locus ranged from 0.55 (DYS531) to 0.96 (DYS385a/b), the haplotype of diversity was 0.9998 for these 24 Y-STR loci. The established 24-plex Y-STR typing system is proved to be stable and efficient in forensic DNA typing.     

Detection of ABO and GM system antigens in the teeth

A series of experiments using agglutination inhibition reaction and material extraction in PBS with pH 7.2 (phosphate-buffer-sedin) has been made to detect antigens of the GM system in the teeth. Parameters of the test material/serum ratio are proposed. In parallel, control specimens of blood were examined. The results of the experiments suggest that detection of Gm antigens in the teeth is feasible. In some cases this may raise an identification level of the expert conclusions. The above technique can be used for investigation of the bones.     

The polymorphism of alpha-amylase]

Individual phenotypes, phenotypic and genetic frequencies of alpha-amylase enzyme were determined by means of population genetic study. The results of this study revealed absence of genetic linkage between alpha-amylase phenotypes, haptoglobins and serum factor G1m(1) of gammaglobulin system (Gm).