非分泌型的基因型与血型物质的分泌研究

阐明非分泌型的基因型与血型物质分泌量和Lewis表型的关系。应用时间决定性荧光免疫测定法（TR－FIA），检测传统的A型Lewis阳性非分泌型个体唾液中H、A及Lewis抗原含量，并以序列特异性PCR，确定其基因型。非分泌型个体唾液中检出了高含量的Lea抗原，其中8例不同程度的检出了少量H、A和Leb抗原，其FUTZ位点为se2／se2纯合型，属Le（a＋b＋）型；1例基因型为se3／se5杂合型，未能检出H、A及Leb抗原，属Le（a＋b－）型。se2是弱分泌基因，se3及se5是非分泌基因。     

唾液中H及H2血型物质的定量研究

用抗H1E3（H1及H2特异性）及抗H224（H2特异性）抗体，以时间决定性荧光免疫测定法对129例个体唾液中的H及H2血型物质进行定量检测。证明在分泌型唾液中含有大量H物质，其主体是H1物质，H2物质的含量为H物质的1／10～1／20。在非分泌型唾液中检出少量H1物而无H2物质，表明H2物质的检出与否可作为区分分泌型与非分泌型的标准，推测唾液中H物质含量的差别，由Se．H基因编码的两种α（1→2）岩藻糖基转移酶活性差异所决定。     

抗Le~a和抗Le~b血清的研制

利用进口抗Le~a、Le~b血清筛选OLe(a+b-)和OLe(a-b+)型人,测定其唾液中Le~a和Le~b型物质含量,择其型物质含量高者唾液,用家兔和山羊免疫,合格后采全血,分离血清,用O、A、B型红细胞吸收、除去种属及α和β等凝集素。再用O型Le非相应型红细胞吸收,精制出含不完全抗体的抗Le~a和抗Le~b血清。该血清对Le相应型红细胞均产生明显凝集反应,而不发生非特异的交叉凝集反应。经过盲测鉴定,17份红细胞的Lewis型测定完全准确。制备的抗Le~a和抗Le~b血清在效价和特异性方面达到了引进的同种血清水平,填补了我国抗Le~a、抗Le~b血清制造的空白。     

人类分泌状态的研究及进展

ABO血型系统属人类的重要遗传标记之一。ABO抗原不仅存在于红细胞膜，也广泛存在于人体的体液和分泌液中。在体液及分泌液中分泌ABH血型物质的人称作分泌型（Seers－tors，See）。A型人分泌A物质，B型人分泌B物质，O型人分泌H物质。所有分泌型人都有H物质的分泌；不分泌ABH血型物质的人称作非分泌型（nonsecretors，uSe）[1]。分泌ABH血型物质，首先是在检测人精液和唾液时被证实[2]。后来的研究结果表明：在人类分泌型个体的消化道（唾液、胃粘膜、胆汁、胎粪）、泌尿生殖道（精液、阴道分泌物、卵巢囊肿液、尿）、呼吸道及乳汁…     

H-抗原缺乏分泌型个体FUT1及FUT2等位基因的分子基础

一例H-缺乏分泌型个体被发现。其血清学表现为：红细胞上无A、B、H抗原；唾液中有B、H抗原分泌；血清中检出了抗A抗体。推测其为类孟买OHmB型。在该个体FUT1等位基因编码区上发现两处单碱基突变（T460C、G1042A）。这两个点变异，将导致两个氨基酸的置换（Y154H、E348K）。同时也破坏了限制性内切酶RsaⅠ和AvaⅠ的作用部位。用PCR－RFLP法可检出此两种变异。用PCR－RFLP法证实，该个体为T460C、G1042A变异的纯合子。在136例随机个体中未能查出上述变异。将该FUT1基因转染COS－7细胞未能检出α2－FUT活性及证实H抗原的表达。该个体的FUT2基因与野生型一致。     

矛盾分泌型精斑检验1例

1案情摘要1995年6月某日晚九时许，周某报称被人强奸．从周某被强奸时所穿的短裤上检出人精斑，笔者用中和法（凝集抑制试验）在斑速处检出B血型物质，确定为B血型精斑，而重大嫌疑人赵某血样呈AB型，似被排除作案可能．然而在对其唾液进行ABO血型检验时，意外发现其唾液中仅检出B型物质，未检出AH物质，与体液ABH物质分泌规律不符，表现为矛盾分泌型．其血型物质与精斑检材中血型物质一致，故不能排除其作案可能．案件经全面调查，最后确认赵某即为本强奸案的犯罪分子，且赵某对作案经过供认不讳．2讨论通常根据体液（唾液、精液等）…     

恒河猴ABO血型的研究

本文应用A型、B型、AB型血清，及抗－A与抗－B单克隆抗体对113只猕猴的ABO血型进行了测试。结果表明：在恒河猴（Macacamulatta）红细胞表面及血清中均未证明有类人凝集原和凝集素存在。但可根据其唾液中类人ABH型物质存在的情况判定该动物的类人血型。本实验检出的血型表型频率分别为：A型17．70％；B型52.21％；AB型20.36％；O型9.73％。该结果与国外报道的血型分布频率有差异。     

人精浆中A型血型物质的分离纯化及理化性质研究

应用凝胶过滤、阴离子高速液相色谱及免疫亲合层析方法成功地从A型分泌型人精浆中分离、纯化了具有A型活性的精浆血型物质（SPBGS－A）。理化分析结果表明：SPBGS－A为表现单一A型活性、分子量不均一的糖蛋白。其分子量约为100KD；等电点在pH2．7～3．5之间；糖含量占72．5％；蛋白质含量占24．2％。共检出了16种氨基酸成分，其中苏氨酸、丝氨酸及脯氨酸的含量约占50％。经与其它水溶性血型物质在化学组成上进行比较，提示其可能具有一定的器官特异性。     

中和法检验人体液斑中H型物质含量在个人识别中的应用4例

中和法检验人体液斑中Ｈ型物质含量在个人识别中的应用４例刘仙洲（河北省顺平县公安局；顺平０７２２５０）Ａ、Ｂ、Ｈ类血型物质不仅存在于红细胞上，在唾液、精斑、阴道分泌液及人体各组织也广泛分布。分泌型人的体液中除分泌与本身相同的型物质外，还可分泌Ｈ型物质，...     

ABO基因分型及其在法医学中的应用

为建立一种ABO血型系统基因分型方法，采用PCR-RFLP技术，成功地将ABO系统区分为AA，AO，AB，OO，BB，BO六种基因型。对240名中国汉族无关个体血样的ABO（基因型频率调查结果表明，6种基因型的频率分布为0．0125～0．3834，符合Hardy－Weinbeng遗传平衡法则（P＞0．1），其DP值为0．8161。家系分析表明，亲代a、b、o基因传递遵守孟德尔遗传规律。对法医学中常见的血痕、混合斑、骨组织及毛发根部等生物样品进行检测，均能准确判定ABO基因型，并可在实际案件鉴定中应用。     

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.     

Lewis blood group determination in bloodstains by planimetric measurement of eluted monoclonal antibodies

Planimetric measurements were employed for reading the results of an elution test to determine Lewis blood groups in dry human bloodstains. In the absorption-elution test, two varieties of indicators were used to detect eluted Lewis antibodies. First, 64 blood-stains aged between 2 to 8 months were tested with glutaraldehyde (GLA)-treated erythrocytes (planimetric hemagglutination assay, PMHA). This method demonstrated that dry stains weighing approximately 0.4 mg (equivalent to 3 microliters of whole blood) were sufficient for detection of Lea or Leb antigen. Results were obtained within 1 h. Then, 37 of these stains were tested with Lewis substance-coated latex particles (planimetric latex agglutination assay, PMLA). The presence of Lea and Leb antigen were detected from dry stains weighing 0.1 mg (equivalent to 1 microL of whole blood) within 3 h. Both these assays are faster and simpler with accuracy than the enzyme-linked immunosorbent assay (ELISA). Latex particles coated with Lewis substance are, in particular, strongly agglutinated and show agglutination patterns more clearly than erythrocytes. The blind tests using these two methods properly classified 7 Le(a + b-) and 23 Le(a-b + ) bloodstains; whereas, 5 Le(a-b-) stains were undetermined by the criteria for these tests. These results indicate the usefulness of the PMHA and PMLA for typing Lewis blood groups from small bloodstains.     

ABH and Lewis antigens of tracheal glands. II. Lewis negative individuals

Immunocytochemical studies were performed on tracheal wall samples embedded in paraffin; the samples were taken at 23 autopsies. In all cases, the red cells had been typed in postmortem serological studies as being Le(a-b-). Blood-group antigens were demonstrated by the indirect immunoperoxidase technique, using monoclonal Anti-A, Anti-B, Anti-Lea and Anti-Leb; H was detected by UEA 1. The secretor characteristics could clearly be diagnosed from the ABH staining pattern of the mucous glands. In 11 cases, the lewis antigen labeling patterns were identical to the group of Lewis-positive individuals. It seems probable, from the statistical point of view, that these 11 individuals were, in fact, Lewis-positive and that the negative serology resulted from deterioration of the cadaver blood samples. The immunocytochemistry was quite different in the remaining 12 cases: (a) secretors (n = 9) were completely negative for Lea, Leb was equally negative in one case, but in the remainder it was detectable within mucous epithelia in minimal amounts and in an atypical granular distribution; (b) nonsecretors (n = 3) reversely exhibited complete negativity for Leb but a minimal staining for Lea. These findings are in harmony with the well established Lewis serology typing of secretions in Lewis negative individuals. Thus, a minimal Lewis antigen biosynthesis and secretion seem to occur in the absence of the Le gene: A alpha-4-L-fucosyltransferase of low activity might be the product of the allele le.     

Studies and observations on Lewis grouping of body fluids and stains

Leb positive individuals may phenotypically express both Lea and Leb in their secreted body fluids. Therefore, the interpretation of a Le(a + ,b-), non-secretor result is dependent on the absence of Leb. This study emphasises the importance of accurate procedure and biased selection of antisera such that Leb is preferentially detected in comparison with Lea. The relationship of the ABO group to the expression of Le is discussed in conjunction with the selection of samples for testing antisera and inclusion as control standards.     

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.     

Determination of the Lewis blood group substances in stains of forensically relevant body fluids

Forensic investigations often demand a clear definition of secretor status. Lewis-typing of secretion stains may help to verify non-secretor results and to identify mixtures of secretions from Le (a-b-) persons and secretors (or non-secretors). Furthermore it gives an additional check on secretor status, determined by ABO-grouping. Few problems may arise, when testing prepared saliva or semen stains. Therefore our interest was focussed on the possibility of Lewis-typing in stains appearing in forensic case work such as cigarette tips, stamps and envelope flaps, semen stains and vaginal swabs, nasal secretion, sweat and urine stains. All stains with the exception of sweat and urine were successfully Lewis-typed. In saliva stains Lewis substances could be determined even after 5 years and in semen stains for at least up to 40 days.     

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.     

A capillary tube method for the Lewis typing of red blood cells

A simple and inexpensive capillary tube procedure, which can be applied in the forensic science laboratory, is described for the detection of the Lewisa (Lea) and Lewisb (Leb) antigens on red blood cells. This procedure will permit approximately 4000 tests to be performed from a single 2-mL bottle of Lewis antiserum.     

Preparation, evaluation of specificity and use of anti-Le monoclonal antibodies

Anti-Le(a) and anti-Le(b) monoclonal antibodies were obtained and attempts at evaluating their epitope specificity have been made. A method for identification of Lewis phenotypes in salivary specimens by dot-I enzyme immunoassay has been developed. Analyses of salivary samples from 72 donors detected donors with phenotypes Le(a-b-) and Le(a+b+), which was confirmed by hemagglutination test.