西安地区人群Hp亚型基因频率分布及其法医学上的应用

触珠蛋白(Haptoglobin简称Hp)是一种存在于人类及其他哺乳动物血清和其他体液中具有遗传多态性的X_2糖蛋白.Smithies首先发现了Hp的遗传多态性,用淀粉凝胶电脉技术将其分为三种表现型.嗣后,Smithies与Connell又将Hp分为六种表现型.进入八十年代,Shibata与Mastumoto等人采用等电聚焦电泳技术将Hp进一步分为16种表现型,证实由Hp~(1f)     

用抗Hpα链抗体的等电聚焦,免疫印染方法进行Hp亚型分型

<正> 人Hp由α和β亚单位组成,其α链本身与Hp的基因型有关。虽然已发表了数篇关于利用等电聚焦——免疫印染方法或双向电泳方法进行Hp亚型分型的文章,但要清楚地确定Hp     

新疆维吾尔族群体结合珠蛋白多态性研究

人的血清结合珠蛋白(Haptoglob in’简称Hp)作为一种具有多态性的遗传标记,已被广泛应用于法医学的物证鉴定和个人识别上,并取得了一定的效果。由于Hp的各种表型的分布频率在各地区和民族中是不相同的,并存在着Hp的异常表型,故进一步研究这方面的遗传规律,是为了更好的在法医鉴定中应用Hp遗传标记。     

人血清Hp、Tf、Gc型的同步检测

人血清Hp、Tf、Gc蛋白是在法医学个人识别及亲子鉴定中应用最为广泛的三种遗传多态性蛋白标记。[1][2][3]本文采用聚丙烯酰胺垂直板状凝胶电泳同时进行Hp、Tf、Gc三种蛋白的分型。经氨基黑、联苯胺分别染色后。Hp、Tf、Gc的电泳图谱清晰、重复性好,整个操作过程仅需6小时。作者还用该方法检测了上海地区人群Hp、Tf、Gc的分布频率。     

用快速高效液相色谱系统分离纯化Gc蛋白

报告Gc蛋白的一种分离方法，为免疫制备抗Gc血清制备抗原。硫酸该分级沉淀出含Gc的人血清组份，BlueSepharoseCL－6B亲和色谱除去含Gc硫酸铸分级沉淀的人血清组份中的白蛋白，快速高效液相色谱系统过Alkyl-SuperoseTMHR5／5疏水色谱柱和MonoQHR5／5离子交换柱。聚丙烯酸胺凝胶解离、非解离电泳证明，Gc蛋白得到了彻底纯化。免疫固定显示：纯化出的蛋白确实是Gc蛋白．为1-1型。快速高效液相色谱为蛋白质的纯化提供了新的技术及仪器手段。     

在Gc电泳图谱上读Hp表型

前言血型特异性成分(Gc)和结合珠蛋白(Hp)是两种血清球蛋白,分别具有三种遗传表型Gc1—1,Gc2-1,Gc 2—2,和Hp1-1,Hp2-1,Hp2-2,按孟德尔遗传定律遗传,是法医学个人识别及亲权鉴定的两个重要遗传标记。一般情况下,检测血清Gc和Hp需要分别配制凝胶、凝胶缓冲     

长沙地区人群Hp型的分布调查

<正> 我们采用聚丙烯酰胺凝胶圆盘电泳法检测长沙地区285名献血员的血清Hp表型及基因频率(表1),并用x~2检验Hp型与ABO血型的关系(表2).结果表明,Hp各型的基因频率分布在国内南方地区极相近,与国外比较有一定种族差异;且Hp遗传与ABO遗传之间无相关性.     

PAG圆盘电泳同步检测人血清Hp、Gc、Tf表型

本文采用不连续ＰＡＧ圆盘电泳法，以邻联甲苯胺和氨基黑染液取代有强致癌作用的联苯胺试剂用于染色，同时检测出Ｈｐ、Ｇｃ、Ｔｆ表型。该法简便快速，安全可靠，经济实用，用于法医学检察实际，可使检案质量和办案效率大为提高。     

毛发的裂解色谱分析

本文应用不同极性的3条大口径毛细管柱,对13人的毛发进行了裂解色谱分析。结果不同人毛发裂解谱图不同,同一人不同部位及同一根毛发的不同裂解谱图未发现明显差异。该文还对裂解色谱的重现性进行了探讨,建立了毛发的裂解色谱方法。     

上海地区中国人Hp血清型的分布和遗传调查

<正> 1955年,Smithies 使用淀粉凝胶电泳技术发现 Haptoglobin(结合珠蛋白,简称 Hp)有多态性,并将它分类为3型,即Ⅰ型,ⅡA 型,ⅡB 型。随后 Smithies 和Walker 调查了18个家系的 Hp 出现频     

The application of immunoblotting to the phenotyping of haptoglobin.

An immunoblotting method for phenotyping haptoglobin in serum and bloodstains has been developed. Haptoglobin isoproteins were separated by polyacrylamide gradient gel electrophoresis and then transferred to nitrocellulose by electroblotting. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm thick gels. Nitrocellulose blots were developed by double antibody enzyme immunoassay. The detection limit for serum and bloodstains was improved 16 times compared to conventional staining using O-tolidine. The method could detect haptoglobin phenotypes from 0.001 microliter of whole blood. This detection limit is approximately 8 times lower than that of group specific-component analysis by immunoblotting.     

Haptoglobin phenotyping from older bloodstains by enzyme immunoassay and haptoglobin phenotypes within a Nebraska Caucasian population

Enzyme immunoassay and Western blotting (electrophoretic) techniques were used to determine haptoglobin (HP) phenotypes from older bloodstains. Serum was collected from liquid blood and the HP phenotypes were determined. Bloodstains were prepared from these specimens and stored at various temperatures for several months. The stains were extracted and applied to gradient polyacrylamide gels. The Western blotting technique was used to achieve the transfer of HP bands from the gels to the nitrocellulose membranes. Enzyme immunoassay with goat anti-HP antiserum and rabbit anti-goat immunoglobulin peroxidase were used to identify the HP bands from the extracted samples. Enzyme immunoassay was found to be clearly more sensitive than o-dianisidine or o-tolidine in detecting HP bands from diluted serum samples. The haptoglobin frequency in a Caucasian population in Nebraska was calculated. The frequencies of Phenotypes 1, 2-1, and 2 were found to be 15.8, 48.4, and 35.8%, respectively.     

Haptoglobin phenotyping by polyacrylamide gel isoelectric focusing and its application to simultaneous typing of serum proteins

A simple isoelectric focusing method for haptoglobin (HP) typing is described. Serum was pretreated first with C. perfringens neuraminidase (CPN) and then with dithiothreitol (DTT). The treated serum was subjected to polyacrylamide gel isoelectric focusing (PAGIF), and the band patterns were detected by immunoblotting. The method could be successfully applied to HP typing of bloodstains as old as 2 months. A slight modification of it enabled HP, complement component C81, and factor I (IF) to be typed simultaneously. The immunoblotting facilitated preservation of HP patterns. Thus, the PAGIF method for HP typing is suitable for routine use in the forensic laboratory.     

A new method for typing haptoglobin in bloodstains using immobilized allo A lectin

Allo A lectin from the beetle, which is beta-D-galactose specific, reacts to haptoglobin but not to hemoglobin. The use of allo A-Sepharose for typing haptoglobin in bloodstains helped eliminate hemoglobin from the bloodstain extract and presented highly resolved haptoglobin patterns by disc gel electrophoresis. This method is simple and rapid for typing haptoglobin in bloodstains and can be easily used in forensic science laboratories.     

An efficient method to eliminate streaking in the electrophoretic analysis of haptoglobin in bloodstains

A bloodstain extraction procedure that improves the analysis of haptoglobin in dried bloodstains has been developed. The streaking of electrophoresis gels caused by deteriorated hemoglobin can be eliminated by incorporating chloroform in the bloodstain extraction procedure. The method is easier to execute than previously published techniques for eliminating the adverse effects of deteriorated hemoglobin on the analysis of haptoglobin. Bloodstains up to two years old were correctly phenotyped in haptoglobin by this method.     

Distributions of genetic markers in United States populations: III. Serum group systems and hemoglobin variants

All published and unpublished population frequency data that could be located for U.S. populations are tabulated and presented for the serum group systems haptoglobin (alpha-chain), group specific component, and transferrin and for the common beta-chain variants of hemoglobin. Results obtained by combining data for comparable racial/ethnic groups are also presented. Some evidence is presented to indicate that the results obtained from the combined data may give better information on frequencies for the U.S. population at large than is obtainable from studies conducted in restricted geographic areas.     

Discontinuous polyacrylamide gel electrophoresis for typing haptoglobin in bloodstains

A routine method is described for obtaining reproducible haptoglobin patterns from bloodstains by discontinuous polyacrylamide gel electrophoresis. By employing a stacking gel, proteins from bloodstain extracts are concentrated into narrow zones, before entering the resolving gel. This effect yields highly resolved haptoglobin patterns. Therefore, laboratories without the specialized equipment and expertise for polyacrylamide gradient gel preparation can still obtain highly resolved haptoglobin patterns from bloodstains.     

Haptoglobin typing in canine bloods

Haptoglobin typing by vertical electrophoresis in a discontinuous polyacrylamide gel was conducted on 47 dog blood samples, of which 19 were from Doberman pinschers, 20 from German shepherds, and 8 from pit bullterriers. Two phenotypes were common in the three breeds and could not be used to differentiate between them. Canine haptoglobin phenotypes were, however, sufficiently different from those of humans to warrant using haptoglobin typing as a method for determining the origin of bloodstains.     

Detection of blood opiates by fluorescence-polarization immunoassay

It was found possible to use fluorescence-polarization immunoassay (FPIA) in screening of opium drugs in blood and blood serum as a preliminary test. The results of the study of blood and serum samples by FPIA and chromato-mass-spectroscopy were compared. As a criterion of assessment of the preliminary study results we propose to use a cutoff point by serial production of the analysed blood samples of the threshold morphine concentrations (20 ng/ml for living subjects and 40 ng/ml--for corpses). The method of opiates screening in blood proposed is characterized by fast conduction, high specificity and precision, small amounts of the expert material.     

Detection of haptoglobin from concentrated urine samples by enzyme immunoassay

The detection of haptoglobin (Hp) from serum and bloodstains is utilized extensively in forensic science laboratories in order to include or exclude possible donors. There is an increasing need to make the same discriminations utilizing genetic markers from urine samples. This paper describes the use of enzyme immunoassay and Western blotting (electrophoretic) techniques to determine Hp phenotypes from concentrated urine samples. Serum and urine specimens were collected from volunteer donors. The serum sample from each donor was typed for Hp. The urine specimens were concentrated 3000-fold from the starting volume of 15 mL to a final volume of 5 microL and applied to the gradient polyacrylamide gels. This procedure allows the separation of Hp samples into the three common phenotypes as well as the other rare variants found in humans. The Western blotting electrophoretic technique was used to achieve the transfer of Hp bands from the gels to the nitrocellulose membranes. Enzyme immunoassay with goat anti-Hp antiserum and rabbit anti-goat immunoglobulin alkaline phosphatase conjugate were used to identify the Hp bands from the concentrated samples. Specimens stored for six months at -22 degrees C were also concentrated and typed successfully. Recent implementation of drug-screening policies has resulted in an increase in the submission of substituted urine specimens. The above procedure can be used to detect an additional genetic marker from urine samples and thus facilitate the identity of the donor.