等电聚焦同步检测血浆类粘蛋白ORM_1和α1-抗胰蛋白酶M亚型

作者应用等电聚焦技术,建立了同步检测血浆类粘蛋白ORM_1亚型和α1-抗胰蛋白酶M亚型的方法。本法累计个人识别机率为0.8464,累计非父排除率为0.3739,是同步电泳分型方法中鉴别效率较高的一种。此法为法医学亲子鉴定提供了一种新手段。     

α_2HS-糖蛋白、α_1-抗胰蛋白酶和GC的同步分型与应用

作者应用等电聚焦技术,建立了同步检测α_2HS-糖蛋白表型,α_1-抗胰蛋白酶亚型和GC亚型的方法。本法累计个人识别机率为0.9701,累计非父排除率为0.5811、是国内外同步电泳分型史鉴别效率最高的方法。在11例亲子鉴定中应用本法取得了满意结果。     

人血痕ORM、Pi、AHSG和GC同步等电聚焦分型

作者在国内外首次建立了血清类粘蛋白、α1－抗胰蛋白酶、α2－HS－糖蛋白和型特异性成份等四种遗传标记同步等电聚焦免疫酶放大分型方法。累计个人识别机率为0．9878，非父排除率为0．6648，是国内外已报道同步电泳分型方法中鉴别能力最高的。本法可对稀释度为1／100的血清进行分型。对于室温保存四周的血痕盯全部正确分型。除Pi外，ORM、AHSG和GC三种遗传标记至少在24周之内皆可正确分型。     

郑州地区汉族人群TFC亚型基因频率调查

我们根据谭明等介绍的超薄层聚丙烯酸胺等电聚焦电泳技术检测TFC亚型的方法，略加改进，对郑州地区214例无亲缘关系的健康人转铁蛋白（TF）亚型的分布进行了调查（表1），其结果用Hardy一Weinberg公式检验，x2=0·4119，df=4．0．975＜p＜0．990，即观测值与期望值吻合度良好。本文调查结果与辽宁、广东、成都等地区的调查结果相比较，无显著性差异（P＞0·25）。根据我们的调查结果，郑州地区汉族人群TFC亚型的非父排除率（EPP）为15·67％，个人识别能力（DP值）为0．5412。郑州地区汉族人群TFC亚型基因频率调查@申成斌$河南省公…     

19个常染色体与4个Y染色体STR复合扩增体系的建立

目的建立19个常染色体STR及Amelogenin和4个Y染色体STR基因座复合扩增体系,并对其效能进行评估。方法用五色荧光标记20＋4Y—STR基因座,建立同步扩增检测体系,用ABI3130XL遗传分析仪对扩增产物进行电泳,GeneMapperID3．2软件进行基因分型;检测体系的灵敏度、均衡性、稳定性、特异性、同一性和稳定性,并观察混合、降解及微量检材的分型情况。结果采用本文体系,DNA模板量在0．05～1．00ng时,分型准确,均衡性、特异性好;混合、降解及微量检材分型正确。该19个常染色体STR基因座的累计个人识别率大于0．999999999,三联体累计非父排除率达0．999999985,Y—STR单倍型多态性为0．592。结论本文建立的复合扩增体系分型准确,稳定,在法医学案件检验及数据库建设等方面有良好的应用前景。     

PGM1和EAP同步等电聚焦电泳分型

采用薄层等电聚焦电泳PAGIF，结合反转显色，同时对EAP及PGM1亚型分型。结果在同一块凝胶板以两个表面分别显现出清晰的同工酶谱带，结果互不干扰。EAP检出AA、BB、BA三种表型PGM1检出十种亚型。PAGIF结合双面反转显色可同时对EAP及PGM1亚型分型，累积个体识别率为0．87，可应用于亲子生定。     

血痕Tf亚型检测及北京地区基因频率分布调查

采用超薄层聚丙烯酰胺凝胶等电聚焦法对血痕Tf亚型进行分型,并调查北京地区223名无血缘关系汉族人群Tf亚型的分布。其基因频率:TfC~10.7354,TfC~20.2377 TfDchi 0.0269。经Hardy-Weinberg吻合度检验,观察值与期望值无显著性差异(ΣX~2=0.9905 df=3 p>0.50)。Tf的个人识别率为0.4020。非父排除率为0.1813。讨论了汉族群体Tf亚型及不同地区,不同人种间Tf分布的情况。到目前为止,室温下保存8个月的血痕仍可进行Tf亚型分型。对实验中质量的控制作了探讨,采用伏时控制电泳条件。     

太原地区汉族人群Tf亚型分布调查

本文采用超薄层等电聚焦电泳方法对太原地区汉族人群进行Tf亚型分型,结果表明:该地人群基因频率为:TfC~1=0.7561,TfC~2=0.2244,TfD=0.0171。非父排除率与个体识别率分别为0.1650和0.5579,属高鉴别类多态系统。本文与其他作者的调查结果进行比较,发现本地区人群Tf亚型分布与高加索人及黑人有显著性差异。     

精斑中磷酸葡萄糖变位酶(PGM_1)及其亚型的电泳分型

本文用淀粉凝胶电泳法和 PAGIEF 对精斑 PGM_1普通型及亚型进行了检测。169份精液斑的 PGM_1分型结果是:PGM_1 1—1 87例;PGM_1 2—1 66例;PGM_1 2—2 16例,其中31例同一个体红细胞及精液 PGM_1分型的结果完全一致。研究了138例不同精子数精斑的 PGM_1型,发现精子数的多少对分型无影响。亚型检测结果与红细胞一样可分10型。     

用PCR-SSP法分析中国辽宁汉族HLA-DRBI基因多态性

应用PCR－SSP方法对辽宁地区159名无关个体进行HLA－DRB1位点基因分型，检出8组等位基因（扩增片段大小为100bp），基因频率范围在0．02201～0．23899。36种可能基因型中检出33种。经x2检验符合Hardy－Weinberg平衡定律。本地区汉族群体的期望杂合度为85％，观察杂合度为83％。个人鉴别机率（DP）为0．94非父排除率（EPP）为66％。本法具有简单、快速、结果可靠的特点，不仅适用于法医学亲子鉴定和个人识别、移植配型，亦可用于相关疾病及人类遗传学研究。     

应用等电聚焦电泳对人精液黄递酶Ⅲ表型分布的研究

应用超薄层聚丙烯酰胺凝胶等电聚焦电泳对武汉地区198例成人精液黄递酶Ⅲ(DIA_3)作了分型。观察到4种表现型,基因频率分别为DIA_3~1=0.7727;DIA_3~2=0.2172;DIA_3~3=0.0101。对保存精斑进行检测证实,室温下保存9周的精斑仍能作DIA_3分型。在聚丙烯酰胺凝胶中引入分离剂HEPES,提高了分辨率。改进后的等电聚焦技术尤适于DIA_3基因产物的分离。文中还调查了我国汉族人群DIA_3频率分布,并与其它群体资料作了比较。     

HLA 分型用于亲子鉴定10例报告

本文报导了10起亲子鉴定案例的 HLA 分型结果,并进行了遗传学分析和讨论。结果提示 HLA 分型是一种比较准确地解决亲子纠纷的方法。     

Esterase D phenotyping of bloodstains and hair roots by low voltage isoelectric focusing

A new isoelectric focusing method is described for phenotyping of esterase D in blood stains and hair roots. It permitted easy and rapid discrimination of six phenotypes determined by ESD*1, ESD*2 and ESD*7. Experiments showed it to be practicable in forensic stain work. In addition, this technique was also usable in phenotyping of ESD 5.     

The use of ultrathin-layer agarose gels for phenotyping erythrocyte acid phosphatase by isoelectric focusing

A method is described for the use of ultrathin-layer agarose gels in phenotyping erythrocyte acid phosphatase (EAP) by isoelectric focusing (IEF). The results obtained using ultrathin-layer agarose gels are shown to be equally reliable and reproducible in comparison to established ultrathin-layer polyacrylamide gels. IEF of EAP on 0.168-mm agarose gels took place in 90 min using the LKB Multiphor system. The technique described allows for both time and cost efficient phenotyping of EAP.     

Studies on the use of isoelectric focusing as a method of phenotyping erythrocyte acid phosphatase

The technique of isoelectric focusing (IEF) in ultra-thin polyacrylamide gels as a method of phenotyping erythrocyte acid phosphatase (EAP) has been applied to a large number of red cell lysates and dried bloodstains. This paper presents the results of this study and discusses some features of the IEF patterns and problems with their interpretation. The IEF patterns of several rare EAP phenotypes are also described. These studies have confirmed that IEF is more sensitive than starch gel electrophoresis as a method of phenotyping EAP in dried bloodstains.     

The application of immunoblotting to the phenotyping of group-specific component

A sensitive immunoblotting method for the routine detection of group-specific component (GC) from fresh serum, and from control and casework bloodstains has been developed. GC phenotypes were separated in a thin layer polyacrylamide gel by isoelectric focusing, transferred to nitrocellulose by a rapid capillary blotting procedure, and detected using a double antibody enzyme immunoassay. This method is capable of phenotyping 8 ng of GC extracted from bloodstains, a four-fold increase in sensitivity when compared to immunofixation and silverstaining. A total of 2424 casework bloodstains have been analysed and GC phenotypes identified in 78% of samples. The method is suitable for use in routine laboratories and is more sensitive than other methods for GC phenotyping of casework bloodstains.     

红细胞酸性磷酸酶(EAP)型的分布及血痕EAP的检出

本文应用琼脂糖凝胶电泳法对辽宁地区213例汉族随机献血员的红细胞酸性磷酸酶(EAP)型进行了检测,其基因频率为EAP~(?)=0.169,EAP~b-0.831。结合文献资料分析了EAP型分布的种族差异,指出了各人种EAP型分布的特点。用琼脂糖凝胶电泳法,对红细胞溶血液EAP型的最小检出量为2μL(2×3mm滤纸条)及0.5μL(直接加样);对血痕EAP型的最小检出量为7.5μL血液制成的检样。本法在4℃保存4周以内的红细胞溶血液和室温保存25天的血痕均能正确判定EAP的型别。     

Routine phenotyping of native and desialyzed alpha 2HS-glycoprotein from blood stains

The application of a polyacrylamide gel isoelectric focusing (PAGIEF) and immunoblotting procedure for the identification of native alpha 2HS-glycoprotein (AHSG) in routine casework blood stains has produced reportable results on 57.2% of samples. This reporting rate is lower than that for group specific component (GC) (83.8%) and phosphoglucomutase (PGM 1) (72.8%) phenotyping of the same samples. Blood stain samples were desialyzed with 1 U/ml neuraminidase, overnight at room temperature prior to PAGIEF in gels containing pharmalyte pH 5-6 and 2.5 M urea. Simple AHSG patterns were developed by immunoblotting. This procedure was five times as sensitive as the native AHSG method and desialyzation was reproducible over a range of incubation times and neuraminidase concentrations. The application of the desialyzed AHSG analysis to routine casework samples has resulted in a significant increase in the number of reportable results (762 reported out of 1027 samples). This reporting rate (74.2%) compares favourably with that for GC (79.1%) and PGH 1 (69.6%) phenotyping of the same samples. The three AHSG alleles (AHSG*1, 2 and 3) are clearly resolved after sample desialyzation and separation in gels containing pharmalyte pH 5-6 and 2.5 M urea. The sensitivity of desialyzed AHSG phenotyping approaches that of GC and this technique is worthy of inclusion in blood stain screening protocols of forensic laboratories in regions where the population has a limited range of rare AHSG alleles.     

Haptoglobin phenotyping of bloodstains by horizontal electrophoresis on a compact polyacrylamide gradient gel

A method is described for phenotyping haptoglobin by horizontal electrophoresis on a small polyacrylamide gradient gel. This method employs the same apparatus used in the separation of many red cell enzyme phenotypes and thereby eliminates the necessity for specialized vertical electrophoresis equipment.