Detection of ABH blood group antigens in the saliva of Koreans and their stability according to storage of saliva samples

The purpose of the present study was to identify salivary molecules carrying the ABH blood group antigens in Koreans and to investigate the changes in these antigens according to processing and storage of saliva samples. Secretor or non-secretor phenotypes and salivary components carrying the ABH antigens were identified in 90 subjects, 30 subjects in each ABO blood group, by SDS-PAGE and immunoblotting. Saliva samples were then obtained from 12 secretors-two males and two females in each ABO blood group and aliquots of both fresh saliva samples and their supernatants after centrifugation were stored at room temperature, 4, -20 and -70 degrees C. The same experiments were performed after 1, 3 and 6 months to investigate changes in the blood group antigens. In all 68 secretors, high-molecular-weight salivary mucin (MG1) was found to be the primary carrier of the ABH antigens. A salivary component of approximately 80 kDa also carried H antigen in seven saliva samples of 22 blood type O secretors. The blood group antigens were better detected in centrifuged samples. In saliva samples preserved at room temperature and 4 degrees C, the blood group antigens were either not detected or detected as degraded molecules. No change was found in the blood group antigens in saliva samples preserved at -20 and -70 degrees C for 6 months.     

Light-microscopic examination of ABH and Lewis antigens in human tracheal and epiglottic glands using the avidin-biotin-peroxidase complex technique

The localization of ABH and Lewis antigens was examined in formalin-fixed, paraffin-embedded human tracheal and epiglottic glands using monoclonal anti A, B, H, Lea and Leb antibodies. The mucous cells of the glands showed reactivity with antibodies corresponding to the respective ABO blood groups of the tissue donors. The mucous cells from one blood group A, Le(a-b-) individual showed no reactivity with any antibodies and those from another blood group A, Le(a-b-) individual showed reactivity only with anti A antibody. In individuals from blood group Le(a + b-) of all ABO groups, the mucous cells reacted exclusively with anti Lea. In blood group O, Le(a-b+) individuals, the mucous cells showed intense reaction with anti H and Leb antibodies and weak to moderate reactivity with anti Lea. In Le(a-b+) individuals of A1, B and A1B blood groups, the mucous cells showed strong reactivity with anti A and/or B antibodies, moderate with anti Leb, weak or no activity with anti Lea and absent with anti H. In blood group A2 Le(a-b+) individuals, the mucous cells stained with anti A were weakly stained or completely unstained with anti H antibody, but cells negative with anti A gave strong positive reactions with anti H antibody.     

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.     

人类精子ABO血型抗原的间接免疫荧光研究

应用间接免疫荧光技术,对40份精子标本进行 ABO 血型抗原检测。A 型人精子上存在 A 抗原;B 型人精子上存在 B 抗原;AB 型人精液中,一部分精子带有 A 抗原,另一部分精子带 B 抗原。各血型人的精子均有 H 抗原。精子血型抗原为本身所固有,并非来源于精浆。不同人的精子血型抗原含量各不相等,与其供体是否为分泌状态或强弱无直接关系。精子 ABO 血型抗原主要存在于精子的颈部和顶体等区域。     

A sequence of tests of minute human blood stains for human origin identification and ABO blood grouping

A series of examinations is presented for human origin identification and ABO blood grouping of doubtful minute human blood stains. A blood-stained thread (0.5 cm in length) was first tested to identify human origin by microprecipitation method and then the ABO blood type was determined by both a modified absorption-elution test and a modified mixed agglutination. In the continuous tests, the maximum limits of positive reactions of the microprecipitation method, the modified absorption-elution test, and the modified mixed agglutination were 1:640, 1:160, and 1:2,560 diluted blood, respectively. A and B agglutinogens were more sensitively determined than H agglutinogen. Hemagglutinogens of blood stains on cotton threads were more easily detected than those of polyester ones.     

混合斑中精斑ABO血型的检验

<正> 1988年,壹岐裕志等报告了用吸附抗α_2-SGP 血清的硝化纤维素膜(NCF)检验混合斑中的精斑 ABO 血型,但耗时。本文作者通过对此方法的改进,采用常彩琴等研制的抗人精特异蛋白血清(anti-human seminalpeculiar protein,ASPP),采用蛋清粘片热解离法检验混合斑中精斑 ABO 血型,耗时短,效果好。现介绍如下。     

ABO tissue antigens of Egyptian mummies

ABO groups were investigated on skin (and muscle), bone and hair specimens from 14 Egyptian mummies dating from the Roman period. Samples were tested by the AE (absorption-elution), MA (mixed agglutination) and HIF (histo-immunofluorescence) methods, in order to evaluate the reliability of each method. For half of the mummies (7) the results were concordant on all samples (3-9 samples for each mummy) with all employed methods, suggesting an unequivocal blood group conclusion. For the other seven mummies there were discordant results with the different methods and interpretation of the results was thus inconclusive. HIF seems to be the most reliable method as specific blood group substances are identified on specific histologic structures. Failure to detect tissular ABO antigens was mainly due to excessive resin impregnation.     

Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+

Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases.     

A, B and H group specific substances in human vitreous humor

The presence of A, B and H group specific substances in vitreous humor taken from 105 human corpses was determined. Good agreement was obtained between these group substances and the ABO blood group. The relationship with the secretor type is discussed.     

The rapid determination of the ABO group from body fluids (or stains) by dot enzyme-linked immunosorbent assay (dot-ELISA) using enzyme-labeled monoclonal antibodies

Using ABH enzyme-labeled monoclonal antibodies, the authors could rapidly detect the ABO group from body fluids and body fluid stains by the dot enzyme-linked immunosorbent assay (dot-ELISA). In this test, the antigen was immobilized on nitrocellulose paper; the entire piece of paper was coated with an appropriate dilution of enzyme-labeled McAb directly against the antigen of interest; and, finally, 3,3'-diaminobenzidine (DAB) substrate solution was added. The site of a positive reaction is clearly visible as a brown spot. We analyzed 521 samples and got satisfactory results. We also analyzed 99 practical case samples by this method and achieved the same results as those obtained by other researchers using other methods. This method is accurate, simple, direct, rapid, and sensitive; it also produces easily observed results, requires no equipment, and can be completed in 30 min. The test proved to be clearly more sensitive for the detection of the ABO blood group in secretor saliva than the conventional hemagglutination inhibition test. Also saliva diluted 10(-4) to 10(-5) and the ABO group of nonsecretor saliva and urine could be easily detected by this method.     

Simultaneous detection of ABO and Secretor-nonsecretor blood groups from forensic biological samples by fragment analysis

This paper describes the simultaneous detection of ABO and Secretor-nonsecretor (SE) blood groups from forensic biological samples by fragment analysis using the ABI PRISM^® 3130 genetic analyzer. The method allows the assay of well-known base changes at three nucleotide positions 261, 796 and 803 on cDNA of the ABO gene, and at 385 and 428 on cDNA of SE gene and a SE pseudo gene, so that reliable group prediction is established by the presence of representative alleles. As a result, simultaneous detection of ABO and SE blood groupings from biological samples was correctly determined by our methods.     

成人肾脏ABO组织血型1-4型糖链H物质的分布

目的观察1型H、2型H及3/4型H糖链在成人肾组织中的分布及其与分泌状态的关系。方法 应用抗ABO抗体及3种糖链特异的单克隆抗H抗体的免疫组织化学方法,检查分泌型与非分泌型个体肾组织中相应抗原物质的分布。结果 在分泌型和非分泌型人的肾远曲小管均表达2型H和3/4型H物质,1型H和3/4型H物质只在分泌型人肾集合管表达,在非分泌型人中不表达。另外集合管的2型H物质的表达与分泌状态无关。结论 人肾组织有ABH物质的表达,不同肾组织细胞表达的H物质结构差异与AB0型分泌状态有关。     

Immunoenzyme demonstration of isoantigens A, B and H in kidney tissue changed by putrefaction

In contrast to blood serology, which usually fails in specimens more than a few days old, immunohistochemistry (PAP technique) provided reliable information on the blood group (ABO) and, in most cases, also the secretor character of 23 kidney specimens stored for months at room temperature. Better results were obtained with monoclonal antibodies than with human sera. In the late stages of decomposition, blood group diagnosis is based on the more decomposition-resistant antigens of the collecting tubular epithelium (in secretors) and the endothelia of the arteriolae medullares rectae and not on the identification of erythrocytic antigens. In addition, a decomposition-resistant epithelial antigen in the distal convoluted tubules (Tc II) is unmasked by autolysis or heterolysis. "Blood group" antigens were frequently detected in bacteria and fungi. These antigens, however, were clearly distinguishable from blood group characters of the tissue. A transient, weak, false-positive reaction with monoclonal anti-B appeared in decomposed Tc II epithelia.     

Immunohistochemical determination of maternal and child blood groups (ABO) in mature placental tissue

Using the indirect immunoperoxidase technique (PAP method), ABO characteristics of mother and child were correctly identified in tissue specimens from 10 mature human placentas. In one case, a weak infantile A reaction was overlooked in the agglutination test but correctly identified by immunohistochemistry. In accordance with the weak expression of ABO characteristics in cord blood, immunohistochemical labeling of infantile erythrocytes with monoclonal and human antibodies, as well as Ulex europeaeus agglutinin I (UEA I), was less pronounced than that of mature erythrocytes. Labeling of the chorionic vessel endothelium, in contrast to that of adult endothelial tissue, was negative with anti-A or anti-B but, regardless of the infantile blood group, pronounced with UEA I. Regular identification of the blood groups was possible in decomposed placental tissue stored at room temperature for 1 week, but not in tissue stored for 2 or more weeks.     

Immunoenzyme determination of ABO and secretor status in paraffin-embedded autopsy material

Using the indirect immunoperoxidase technique (PAP method), A, B, and H antigens were identified on formaldehyde-fixed, paraffin-embedded kidney tissue from 100 autopsies. Comparison with the serologic findings showed all our blood group determinations to be correct. The labeling of the collecting tubules was evaluated as characteristic of the secretor. The secretor status determined according to this parameter was unequivocally confirmed by the Lewis constellation in 78 of 82 cases; group Le(a-b-) could be differentiated with immunohistochemical methods in secretors and nonsecretors. Determination of ABO blood groups and secretion behavior with immunohistochemical methods was correct even in those cases where classic serology failed due to hemolysis and decomposition. Immunohistologic results obtained with monoclonal antibodies were better than those obtained with human sera.     

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.     

Lectin-histochemical detection of degenerative glycoconjugate deposits in human brain

Several lectins were used to study the localization of glycoconjugates in brain of elderly people and patients with Alzheimer type dementia (ATD) and Down's syndrome (DS). Five kinds of degenerated or deposited materials stained clearly by lectins specific to GalNAC, Gal, Fuc, and/or Man were recognized much in ATD and DS, less in elderly peoples, in addition to the binding of the lectins to neurons. (i) Round shape deposits called corpora amylacea (CA) which consisted of various sizes of round material, existed mainly on the surface of cerebral cortex and some in white matter of the brain. They were colored by Alcian blue (AB), Aldehyde fucsin (AF) and periodic acid shiff (PAS) and weakly by Hematoxylin (H), but not by Eosin (B). They showed clear reactivity with lectins specific to GalNAC, Gal, Fuc and Gal-GalNAC. (ii) Amorphous and variform amyloid deposits existed around blood vessels in the white matter were stained by thioflavin and lectins specific to GalNAC, Gal and Fuc, but not with Man specific lectins and PAS, AB, AF and HE. (iii) Another kind of amyloid deposits which showed a similar characteristic to the previous one and were recognized mainly in white matter and independent blood vessels. These deposits were stained by thioflavin but not by PAS, AB, AF and HE and showed good reactivity with lectins specific to GalNAC, Gal, Fuc, Gal-GalNAC, Gal-GIcNAc and Man. The reactivity with lectins specific to Gal, Fuc, and Man was seen in senile plaques (iv) and neurofibrillary tangles (v). Although at present we are unable to explain the origin of these deposits, it is clear from this study that the glycoconjugates form an integral part of the degeneration in the brain. The lectin staining with GS-I is useful in the forensic pathology to diagnose brain disorders at postmortem examination, since these lectin were able to detect five types of degeneration changes and/or deposits.     

PCR－RFLP技术鉴定ABO基因型的法医学应用研究

目的建立PCR－RFLP、非变性PAG胶垂直电泳和银染技术进行ABO基因分型的方法体系,并对200名广东汉族人群ABO基因型频率进行了调查。方法用Chelex－100和酚、氯仿抽提法处理样本,PCR扩增后用非变性聚丙烯酰胺凝胶垂直电泳和银染技术检测分型。结果ABO位点特异性扩增片段长度为175bp～210bp,6种基因型频率分布为0.0250～0.4300,杂合度H值为0.5162,个体识别力DP值为0.7111。结论该方法可成功运用于血液、血痕、精斑、毛发、骨组织和混合斑等检材的个体识别及亲权鉴定的检验。     

ABO blood grouping of hairs using an avidin-biotin-peroxidase complex technique

Fifty-nine hair specimens obtained from human autopsies and volunteers were used for the determination of ABO blood group substances using the ABC (Avidin-Biotin Complex) technique. Positive staining for A, B and H blood group substances was detected only in the medulla of the hairs. Blood group antigens could not be detected in seven hair specimens because they possessed no medulla. Forty-seven specimens obtained from fresh cadavers and volunteers gave the correct results corresponding to the blood group of the donor, but some specimens from individuals of blood group A2, Le(a + b-) showed weak reaction with anti-A and strong reaction with anti-H. The staining intensity with anti-B and -H in some individuals of blood group AB was stronger than with anti-A serum. Five hair specimens obtained from decomposed bodies were also examined. The blood group antigens could be specifically detected in hairs obtained from two exhumed and one putrid body, but no positive reactions were obtained from two cases of drowning where the bodies had been in the sea for about 6 months. In a blind trial, hair specimens from 28 individuals were also examined. Twenty-two specimens which possessed a medulla gave the correct result. Six specimens gave no result because they possessed no medulla.     

Minisequencing-based genotyping of Duffy and ABO blood groups for forensic purposes

Duffy and ABO blood group genetic polymorphisms were studied by minisequencing analysis of single-nucleotide polymorphisms (SNPs) at nucleotide positions--33, 125, 265, and 298 of the Duffy gene and at nucleotide positions-261, 297, 467, 646, and 703 of the ABO gene. In an Italian population sample, we found four alleles and seven genotypes for the Duffy and six alleles and 16 genotypes for the ABO systems. The lower limit for reproducible results was 200 pg DNA, with a range of up to 10 ng and an optimum at 1 ng. All of the 16 analyzed inclusive paternity tests were also consistent with parentage and two out of four inconsistencies with parentage cases were excluded by one or more SNPs. Although Duffy and ABO SNP typing show lower informativeness than most current forensic tests, their robustness, the limited population distribution of FY* Fy type, and the sensitivity of the minisequencing technology suggest that these markers can be useful in selected forensic applications.