Plasminogen (PLG): a useful genetic marker for paternity examinations

The genetically determined polymorphism of plasminogen (PLG) was analyzed by isoelectric focusing on polyacrylamide gels. For analysis neuraminidase-pretreated sera were used. PLG was developed functionally by activation with urokinase and subsequent lysis of casein in an agar overlay. In a random sample of 957 unrelated healthy individuals from Southern Germany, three common phenotypes, PLG1, 2-1, and 2, and five rare variants were found. The allele frequencies were: PLG*1 = 0.7174, PLG*2 = 0.2780, and PLG*Var = 0.0046. The theoretical exclusion rate in cases of disputed paternity is 16.5%.     

Haptoglobin subtyping by isoelectric focusing in ultrathin-layer polyacrylamide gels. Population genetic data for Hanover and Lower Saxony

This report describes a method for subtyping haptoglobin by means of isoelectric focusing in 0.2-mm ultrathin-layer polyacrylamide gels. Haptoglobin (Hp) is purified by ion-exchange chromatography and reduced. The well-known advantages of ultrathin-layer gels combine high isoelectrophoretic resolution of the Hp subtypes with less demands for time and material and make sequential visualization by fixation and protein staining possible. The distribution of the Hp subtypes in 1500 unrelated adults from Hanover and Lower Saxony is presented. Allelic frequencies are calculated to be: Hp*2FF = 0.0030; *2FS = 0.5620; *2SS = 0.0290; *1F = 0.1537; *1S = 0.2523. Segregation analysis for 68 matings shows an autosomal codominant mode of transmission in all cases. For the population investigated the chance of isolated paternity exclusion with the subtyped Hp system amounts to 33.91%.     

HLA-A位点DNA分型及其法医学应用

应用聚合酶链反应0寡核苷酸探针（PCR－SSO）斑点杂交技术，对222名辽宁地区汉族人群无关个体进行HLA－A基因检测，研究中国辽宁地区汉族群体的HLA－A座位基因分布状况。共检出HLA－A等位基因24个，其中以等位基因HLA－A0201最为常见，频率为0．2635；依次是2402101和1101，等位基因频率分别为0．1847和0．1262.理论杂合度为87％，个人鉴别机率为92％，非父排除率为73．3％。在中国辽宁汉族中检出73种基因型，对观察值和期望值进行X2检验，符合Hardy－Weinberg平衡定律（x2＝6．28，df＝9，0．5＜P＜0．75）。家系分析结果表明按照孟德尔方式遗传。提出的中国辽宁汉族HLA－A等位基因的遗传基因情况，可用于法医学个人识别和亲子鉴定。人类学，HLA相关疾病，及器官移植研究。     

Gc subtypes: demonstration using isoelectric focusing. Study of population and formal genetics

Gc-subtypes were determined by isoelectric focusing and immunfixation on samples from 492 unrelated blood donors from Berlin. The frequency of the three genes was found to be GcIF = 0.1270, GcIS = 0.6006, Gc2 = 0.2724. Analysis of 78 parents with 190 children did not show deviations from the expected mode of inheritance. Investigation of the adults from paternity cases and of their children on the other hand obtained similar results. No rare alleles were observed.     

Polymorphism of plasminogen in Tuscany (Italy)

The distribution of PLG phenotypes in the population of Tuscany (Central Italy) has been investigated by means of isoelectric focusing followed by immunofixation of desialyzed sera. In a random sample of 383 unrelated healthy blood donors registered at the Hospital of Pisa, three common phenotypes, PLG A, A-B, and B, and two rare variants were found. The allele frequencies calculated in our study were: PLG*A = 0.6749, PLG*B = 0.3225, and PLG*rare = 0.0026. The theoretical exclusion rate in cases of disputed paternity is 17.42%.     

吉林地区朝鲜族唾液酸性富含脯氨酸蛋白遗传多态性的研究

应用聚丙烯酸胺凝胶等电聚焦技术，调查了吉林地区226名朝鲜族个体唾液酸性富含脯氨酸蛋白二位点上共6种等位基因频率的分布：PRH1*1为0．0331，PRH1*2为0．2124，PRH1*4为0．7477，PRH1*6为0．0068；PRH2*1为0．7544，PRH2*2为0．2456。按Hardy－Weinberg法则进行吻合度检验，其观察值和期望值一致，并对吉林地区朝鲜族与其它地区人群酸性富含脯氨酸蛋白等位基因的差异性做了比较。PRH1和PRH2在吉林延边地区朝鲜族的个人识别能力分别为0.58和0．53，两者总鉴别机率为0．80；PRH1和PRH2的非父排除率为0．1875和0．1510，两者总非父排除率为0．3102。     

Polymorphism of the human phosphoglycolate phosphatase in Northrhine-Westphalia (F.R.G.) and its application to paternity testing

Blood samples from 563 unrelated German and from 110 Turkish individuals living in the Düsseldorf area were studied for phosphoglycolate phosphatase polymorphism. The distribution of the observed phenotypes in the population genetic study did not diverge from the expected values according to Hardy-Weinberg law. In our series, the gene frequencies were calculated as follows: (a) Germans: PGP1 = 0.851, PGP2 = 0.118, PGP3 = 0.031, (b) Turks: PGP1 = 0.973, PGP2 = 0.018, PGP3 = 0.009. The assumed autosomal codominant mode of inheritance was confirmed by the examination of 109 mother-child pairs and by analysis of 70 cases of disputed paternity. The plausibility to exclude German non-fathers from paternity is 12.78%.     

Transferrin polymorphism detected in human urine using isoelectric focusing followed by immunoblotting

Genetic polymorphism of transferrin (TF) was revealed in human urine by isoelectric focusing and immunoblotting on thin-layer polyacrylamide gels. Using this technique more than 300 urine samples were examined, and correct TF typing from a small volume of urine (approx. 0.5 ml) was achieved, in comparison with the results of direct grouping for plasma. Three common phenotypes, TF C1, C2-1 and C2, were differentiated. In addition, the rare types TF C1D, C2D, and C1B were observed. The frequencies of the TF alleles in our samples were found to be: TF*C1 = 0.7265, TF*C2 = 0.2624, TF*D = 0.0083 and TF*B = 0.0028.     

EsD*5 gene frequency in Tuscany (Italy)

The distribution of the human red cell esterase D (EsD) "extended" polymorphism in a population sample from Tuscany (Italy) was studied using agarose gel isoelectric focusing. The estimated gene frequencies were: EsD*1 0.864, EsD*2 0.115, EsD*5 0.021. The EsD*5 allele frequency is very similar to those reported for other European populations. The "extension" of the EsD polymorphism may prove to be useful in paternity testing.     

Genetic polymorphism of desialyzed alpha 2 HS-glycoprotein by ultrathin isoelectric focusing

The genetically determined polymorphism of alpha 2 HS-glycoprotein was analyzed by immunoblotting ultrathin-layer polyacrylamide gel isoelectric focusing in the pH range 4-6.5 and neuraminidase pretreated sera. In a Libyan population sample from Tripoli (n = 110) three common phenotypes, alpha 2 HSG 1-1, 2-1, and 2-2, were observed. The allele frequencies were alpha 2 HSG1 = 0.8364 and alpha 2 HSG2 = 0.1636. The theoretical exclusion rate in cases of disputed paternity is 11.8%.     

亲子鉴定中STR基因座的基因突变分析

目的探讨Identifiler^TM荧光标记复合扩增试剂盒15个STR基因座在亲子鉴定中的基因突变特点。方法应用Identifiler^TM荧光标记复合扩增试剂盒检测676例亲子鉴定案，对其中1～2个突变基因座加做HLA等位基因检测或Y—STR基因座检测。结果在认定亲子关系的676例中，观察1304次减数分裂，Identifiler^TM荧光标记复合扩增试剂盒中的15个基因座确定19例突变，其中D18S51基因座4例，D2S1338基因座3例，D8S1179、D16S539、vWA、D7S820、D13S317基因座各2例，D5S818和TH01基因座各1例，D21S11、FGA、D3S1358、D19S433、TPOX、CSF1P0基因座未见突变；一步突变的17例，二步突变的为1例，四步突变的1例；1个基因座发生突变的18例，2个基因座同时发生基因突变的为1例；突变来自父亲与来自母亲的比例为13：2，4例来源不能确定。结论用Identifiler^TM荧光标记复合扩增试剂盒检测到1—2个基因座发生突变，须增加对其它遗传标记的检测。     

C6 phenotyping in sera and bloodstains by isoelectric focusing followed by electroimmunoblotting

The genetic polymorphism of C6 was investigated in 329 unrelated Japanese individuals using isoelectric focusing in polyacrylamide gels followed by an electroimmunoblotting technique. Besides six common phenotypes C6 A, AB, B, AB2, BB2 and B2, six rare variants were observed. The allele frequencies were: C6*A = 0.4422, C6*B = 0.4757, C6*B2 = 0.0714, C6*A3 = 0.0015, C6*M1 = 0.0046 and C6*B3 = 0.0046. The population data confirmed that the C6*B2 allele is the third common allele characterizing Japanese. The present electroimmunoblotting technique was applied to demonstrate C6 types in dried bloodstains. The C6 types were determined from bloodstains stored at 4 degrees C for up to 10 weeks, at room temperature for up to 2 weeks and at 37 degrees C for up to 4 days. The results show that this component system offers a new powerful means for the medico-legal grouping of bloodstains.     

Further evidence of a silent plasminogen (PLG) allele in two paternity cases

Routine paternity testing has yielded two different cases of an apparent inverse homozygosity in the plasminogen (PLG) system. In one case, the child presented the phenotype PLG A and his putative father the type PLG B. The alleged father could not be excluded from the paternity in 25 additional blood group marker systems (biostatistical probability of paternity W greater than 99.75%). In the other case an incompatibility was found in a mother- child pair. Analysis of PLG was carried out by isoelectric focusing on neuraminidase-treated sera. In both cases the immunologic and functional detection showed weaker banding pattern of the affected PLG types. The assumption of a silent allele in the PLG system was confirmed by quantitative investigations. The allele frequency of PLG*Q0 in the South German population was estimated to be 0.0013. In the same sample the variant PLG A3 has been shown to be polymorphic.     

DNA数据库9个STR基因座比中认定亲权的可靠性分析

目的探讨Profiler plus试剂盒9个常染色体STR基因座用于DNA数据库中双亲亲缘关系比中结果认定亲权的可靠性。方法在DNA数据库中搜集无关个体与已知母子（或父子）在9个STR基因座不排除亲缘关系的比中记录54例，组成54例假定的三联体家庭，计算其PI值与RCP值；应用Identifiler试剂盒加做到15个常染色体STR基因座，观察其排除情况。结果在54例假定三联体中PI值最低为178．598597（RCP=99．443203％），最高为97318．085812（RCP=99．998972％）。加做到15个STR基因座后，54例假定三联体中每个三联体至少出现2个基因座排除，最多5个基因座出现排除的现象，平均排除基因座数为3．52个。结论Profilerplus试剂盒9个STR基因座用于亲缘关系鉴定可能出现错误结论；单纯利用RCP值来认定亲缘关系是不安全的；建议应用16个或更多的基因座建设DNA数据库和进行亲缘关系判定。     

Genetic polymorphisms of orosomucoid ORM1 and ORM2 in a Japanese population: occurrence of new ORM1 alleles

Genetic polymorphisms of orosomucoid ORM1 and ORM2 in a Japanese population from northern Japan were investigated using isoelectric focusing (IEF) in ultrathin layer polyacrylamide gels containing Triton X-100 and immunofixation. Nine ORM1 phenotypes which are determined by four common and one rare alleles were observed. Two of the identified alleles at this locus were considered to be new. The ORM2 pattern was classified into 14 phenotypes as products of one common and two variant alleles. The estimated allele frequencies were ORM1*1 = 0.668, ORM1*2 = 0.170, ORM1*2.1 = 0.136, ORM1*5.2 = 0.022 and ORM1*7 = 0.004; ORM2*1 = 0.972, ORM2*3 = 0.006 and ORM2*6 = 0.022.     

Red cell phosphoglucomutase (PGM1) subtypes in Egyptians

The polymorphism of the human red cell phosphoglucomutase 1 (PGM1) in samples from Egyptians (n = 134) was investigated using isoelectric focusing in thin-layer polyacrylamide gel. In the studied population samples nine common phenotypes were observed, and the calculated frequencies for the genes PGM1+1, PGM1-1, PGM2+1 and PGM2-1 were 0.6381, 0.0821, 0.2201 and 0.0597, respectively. The observed and expected phenotypes provide a good fit to Hardy-Weinberg equilibrium. The four alleles system will increase the probability of excluding a man falsely accused of paternity to 30% as compared with 16% if the two alleles system is used.     

Isoelectric focusing studies on the PGM1 subtypes in the northern Japanese population

The distribution of the human red cell phosphoglucomutase (PGM1) subtypes in samples from Japanese population (n = 277) living in the Miyagi Prefecture, the northern part of Japan, was investigated by applying the thinlayer polyacrylamide gel isoelectric focusing. In our population sample all the ten common phenotypes were demonstrated, and the estimated allele frequencies for the genes PGM1+1, PGM1-1, PGM2+1, and PGM2-1 were 0.671, 0.107, 0.161, and 0.061, respectively. Family studies (n = 40) indicated an autosomal codominant inheritance and confirmed the four alleles. The new system will increase the probability of exclusion in paternity cases among Japanese to 29.4% compared with 14.3% if the two allele system is used.     

Genetic polymorphism of the B subunit of coagulation factor XIII in Libyans: occurrence of a fourth common allele, FXIIIB*6

FXIIIB phenotypes were determined in neuraminidase-pretreated serum samples by using isoelectric focusing in ultrathin-layer polyacrylamide gels containing 1 M urea and subsequent immunoblotting. In a Libyan population sample from Tripoli, (n = 108) nine different phenotypes as products of four common alleles were recognized, with frequencies as follows: FXIIIB*1 = 0.6574, FXIIIB*2 = 0.2454, FXIIIB*3 = 0.0741 and FXIIIB*6 = 0.0231. It is suggested that FXIIIB*6 is the fourth common allele of the FXIIIB system in this population.     

HIV-1 prevalence in deaths caused by drug overdose in various large cities of West Germany and West Berlin between 1985 and 1988

The development of the HIV-1-prevalence among drug deaths (n = 753) in several German cities (West Berlin, Frankfurt, Munich, Hamburg, Bremen, Cologne and Stuttgart) from 1985 to 1988 was evaluated; in 1988 43% of 674 deceased drug addicts were examined. The regional prevalence rate was between 15 and 25%; only in Berlin 42% of the drug deaths were HIV-infected (cumulated data of all cities over the 4-year-period: 26%). There was no uniform or steady regional development of HIV-1-prevalence in the different cities. The ratio men/women among drug deaths was 3:1. The HIV-1-prevalence among males was 22%, among females 40%. HIV-infected individuals were 2-3 years older than seronegatives. Predictions concerning the trend of prevalence rates are not possible up to now. Continuous monitoring of the HIV-status of drug deaths seems to be a worthful method to evaluate the spread of this disease among the risk group of intravenous drug addicts.     

Transferrin (Tf) polymorphism: An analysis by isoelectric focusing

The polymorphism of the transferrin (Tf) system was studied in a total of 300 unrelated Japanese individuals from Miyagi prefecture, the northern part of Japan, using isoelectric focusing in thin-layer polyacrylamide gel. In our population samples three common phenotypes and nine variants were observed. The calculated allele frequencies were Tf^C₁ = 0.773, Tf^C₂ = 0.212, Tf^D_chi (Chinese) = 0.008, and the combined frequencies for the Tf^B_var (variants) = 0.007. Family data (n = 44) were in accordance with an autosomal codominant fashion of inheritance. The use of isoelectric focusing procedure among Japanese will raise the probability of excluding a man falsely accused of paternity to 15.8% as compared with 1.5% when the conventional electrophoretic methods are used.