The phenotypic frequencies of group specific component and alpha-2-HS-glycoprotein in three ethnic groups. The use of these proteins as racial markers in forensic biology

The phenotypic frequencies of group-specific component (Gc) and alpha-2-HS-glycoprotein (A2HS) were determined in White European, Asian and Afro-Caribbean populations. Typical allele frequencies were observed for Gc, with Gc 1S being the major allele for the first two groups and Gc 1F being the major allele for Afro-Caribbeans. For all groups the dominant A2HS allele was A2HS 1, although Asians had a significantly higher proportion of this allele than the White Europeans. Gc and A2HS either singly or in combination with other blood grouping systems provide good discriminating potential. The A2HS 10 allele was detected with a very low frequency in the White European group (A2HS 10 = 0.0013) and was not detected in the Asian group, while the Afro-Caribbean group had a relatively high frequency of this allele (A2HS 10 = 0.0966). The different distribution of the Gc 1F and A2HS 10 alleles in White Europeans and Asians compared with Afro-Caribbeans, can be used to determine the likelihood of blood coming from an Afro-Caribbean.     

Isoelectrofocusing in the study of the BfF system: existence of two common subtypes of the BfF allele in a Japanese population

The Bf gene frequencies including BfF' allele and BfF' allele in a Japanese population were studied by using the PAGIF method. The results showed the Bf gene frequencies: BfF' allele = 0.0778, BfF' allele = 0.1007 and BfS allele = 0.8215.     

Genetic study of red cell esterase D polymorphism by ultrathin layer isoelectric focusing. Distribution in the Veneto population (Italy)

Human red cell Esterase D (EsD) was analyzed by isoelectric focusing (IEF) on ultrathin-layer polyacrylamide gel with a pH range of 5.0-6.0. Hemolysates were treated with Dithiothreitol to avoid loss of activity and change of the isozyme patterns by in vitro storage effects. In our sample of 951 unrelated persons from Veneto, seven different phenotypes were observed. The following allele frequencies were calculated: EsD1 = 0.8476, EsD2 = 0.1336, EsD5 = 0.0178, and EsDV = 0.0010.     

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.     

STR primer concordance study.

Over 1500 population database samples comprising African Americans, Caucasians, Hispanics, Native Americans, Chamorros and Filipinos were typed using the PowerPlex 16 and the Profiler Plus/COfiler kits. Except for the D8S1179 locus in Chamorros and Filipinos from Guam, there were eight examples in which a typing difference due to allele dropout was observed. At the D8S1179 locus in the population samples from Guam, there were 13 examples of allele dropout observed when using the Profiler Plus kit. The data support that the primers used in the PowerPlex 16, Profiler Plus, and COfiler kits are reliable for typing reference samples that are for use in CODIS. In addition, allele frequency databases have been established for the STR loci Penta D and Penta E. Both loci are highly polymorphic.     

PCR-based DNA analysis of vWA31A,TH01, F13A01, FESFPS,TPOX and CSF1PO

The allele and genotype frequencies of 6 tetranucleotide STR loci were investigated in a sample of 132 unrelated individuals from Chinese Han population. The PCR products were analyzed on 6% denaturing PAGE and detected using fluorescently labeled primers in an automated 377 sequencer(PE). All loci meet Hardy-Weinberg equilibrium. There was no random association of alleles among the 6 loci. The allele frequencies were compared with other population databases. Except locus vWA31A, the observed heterozygosity at other 5 loci was significantly lower than that reported in Caucasian and Black population studies. The calculated DP = 0.99999, PE = 0.9708, pM = 1.059 x 10(-5). The allelic frequency data can be used in forensic identification and paternity testing.     

Allele and genotype frequencies for the STR locus D18S51 in a western German population

Genetic marker typing based on DNA amplification by the polymerase chain reaction (PCR) increasingly is being employed in forensic casework and for paternity testing. Allele frequencies were determined using PCR for 102 unrelated Germans (Rhine area) for the locus D18S51. Twelve alleles were observed, with frequencies ranging from 0.005 (allele 11) to 0.191 (allele 14). The observed heterozygosity was 0.867, and the power of discrimination was 0.968. There was no deviation from expectations under Hardy-Weinberg assumptions (P = 0.451).     

Phosphoglucomutase subtyping in a south Polish population

Phosphoglucomutase (PGM1) subtypes in South Polish population were examined by thin-layer polyacrylamide gel isoelectrofocusing (pH 5-7) using fresh hemolysates from 460 unrelated adults. The allele frequencies in Polish population are as follows: PGM1+1 = 0.6402, PGM1-1 = 0.1185, PGM2+1 = 0.1880, PGM2-1 = 0.0533.     

The potential contribution of MVR-PCR to paternity probabilities in a case lacking a mother.

Minisatellite variant repeat (MVR) mapping using the polymerase chain reaction (PCR) was applied to a paternity case lacking a mother to evaluate the paternity probability. After three flanking polymorphic sites at each of MS31A and MS32 loci were investigated from the child and alleged father, allele-specific MVR-PCR was performed using genomic DNA. It was confirmed that one allele in the child was identical to that in the alleged father at both loci. Mapped allele codes were compared with allele structures established from population surveys. No perfect matches were found although some motifs were shared with other Japanese alleles. The paternity index and probability of paternity exclusion at these two MVR loci were then estimated, establishing the power of MVR-PCR even in paternity cases lacking a mother.     

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.     

p33.4位点扩增片段长度多态性及其法医学应用的研究

以聚合酶链反应（PCR）、聚丙烯酰胺凝胶垂直电泳和银染法对中国100名无关个体小卫星区域p33．4位点的扩增片段长度多态性（Amp－FLPs）进行了研究，检出了8个等位基因。通过BIOTRAC系统进行数据处理．各等位基因重复单位的数目分别为7、10到15，其中在13～14之间发现一差值不足一个重复单位长度的罕见等位基因。片段长度分布于603～1115bP之间，基因频率分布于0.5～33．5％间，杂合度为64％，DP值为84．5％。对5个家庭25名相关个体进行分析，符合孟德尔遗传定律；对同一个体不同组织的DNA进行P33.4位点的分型研究表明，该技术适宜于法医物证检验。此外，本研究以Chelex处理不同检材制备DNA模板用于扩增，率先建立了比常规方法更快、更简便、更为实用的检验方法。     

用FLDAS-PCR和PCR-RFLP检测汉族人群GPT的多态性

目的 建立片段长度差异等位基因特异性PCR(FLDAS-PCR)检测GPT基因型的新方法,并调查武汉地区汉族人群GPT多态性分布。方法 应用FLDAS-PCR、PCR-RFLP分别对武汉地区248例汉族个体进行GPT基因型分型。结果 FLDAS-PCR与PCR-RFLP分型结果完全一致,GPT的3种基因型分型明确,基因频率GPT*1=0.5423,GPT*2=0.4577,基因型分布符合Hardy-Weinberg平衡。结论 FLDAS-PCR和PCR-RFSP分型GPT在法医学鉴定中有实用价值。     

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.     

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.     

Study on the application of parent-of-origin specific DNA methylation markers to forensic genetics

In paternity test, especially in motherless cases, the allele inherited from father (obligatory gene, OG) often cannot be determined. The paternity exclusion probability (PE) of a genetic marker is reduced considerably. Therefore, it is necessary to develop a new technique, by which the parental origin of alleles can be determined without genealogical analysis. In this paper, we explored the possibility of using parent-of-origin specific DNA methylation markers to determine the parental origin of alleles, choosing the imprinted single nucleotide polymorphism (SNP) locus rs220028 (A/G) as a model system. We typed the SNP by mutagenically separated PCR (MS-PCR). The frequencies of alleles were A = 0.5085, G = 0.4915; the unbiased heterozygosity was 0.5020. In order to discriminate between the maternal allele and paternal allele, post-digestion MS-PCR, a novel PCR based methylation analysis and SNP typing technique was developed and performed on 18 heterozygous children, and the methylated maternal allele was detected specifically. As a pilot study on the use of epigenetic markers in forensic genetics, our results demonstrated the feasibility of using parent-of-origin specific DNA methylation markers to determine the parental origin of alleles.     

精浆r—谷氨酰转移酶多态性研究

本次实验采用聚丙烯酰胺阶段梯度凝胶电泳方法,对武汉地区135名无关男性精液γ—GT进行了遗传多态性调查,根据γ—GT的电泳差异,分为1型、2型、2—1型三种常见表现型,1型为靠近正极的单一带,2型为靠近负极的单一带,2—1型为1型带和2型带组成的杂合双带。表现型频率分别为0.156,0.356,0.488,基因频率为γ—GT~1=0.400,γ—GT~2=0.600(P>0.5)。室温下保存8周的精斑,也能检测出较清晰的谱带。本次实验还探讨了单纯阴道分泌物及精液、阴道分泌物混合斑进行γ—GT分型的可能性。     

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%.     

Spanish population data on the loci D13S317, D7S820, and D16S539 generated using silver staining (SilverSTR III Multiplex).

A set of 212 samples from unrelated Spanish Caucasians living in Andalucia (southern Spain) were analyzed with a new commercially-available kit for multiplex amplification of 3 STR loci (D13S137, D7S820, and D16S539), manual denaturing polyacrylamide gel electrophoresis and silver staining. These three loci are of special interest for the forensic community since they are a part of the 13 CODIS-core STR loci. The results show that the loci D13S317 and D16S539 meet Hardy-Weinberg expectations (HWE), but the locus D7S820 did not meet HWE (p = 0.003). However, there was no detectable departures from independence (i.e., linkage disequilibrium) between any pair-wise combination of loci. The D7S820 data were further investigated. The excess homozygosity was due to an excess of D7S820 10, 10 homozygotes. To determine if the allele frequency data are meaningful and can be applied to forensic identity cases, the Spanish D7S820 allele frequency data were compared with four other Caucasian sample populations. The D7S820 allele frequencies were statistically similar; thus, the results support that the allele frequency data can be used reliably for estimating DNA profile frequencies.     

Allelic distribution of four tetranucleotide repeat loci (D3S1358, D18S51, D19S253, and FGA) in a population from Porto (North Portugal)

Allele frequencies for four short tandem repeat loci were determined in a population sample from Porto (North Portugal), using the polymerase chain reaction (PCR), in order to investigate possible genetic differences between populations from the center and north of Portugal. After denaturing PAGE electrophoresis, nine alleles were identified for D3S1358 (n = 256), 13 alleles for D18S51 (n = 235), 10 alleles for D19S253 (n = 238), and 15 alleles for FGA (n = 181). No deviations from Hardy-Weinberg equilibrium were found. The allele frequencies observed are similar to those of the Portuguese population compared except for the D3S1358 system.     

α-纤维蛋白原基因多态性与肺动脉栓塞相关性初探

目的探讨α-纤维蛋白原Thr312Ala基因的多态性分布与肺动脉栓塞的相关性。方法选取尸检已确诊的肺动脉栓塞病例的甲醛固定心肌组织14例和正常健康人群外周血56例,采用改良酚-氯仿法和Chelex100法分别提取心肌组织和外周静脉血基因组DNA,用PCR-RFLP法检测α-纤维蛋白原基因Thr312Ala多态性,并分析其与肺动脉栓塞的关联性。结果α-纤维蛋白原Thr312Ala基因的各基因型在肺动脉栓塞组与对照组比较差异具有显著性意义（P〈0.05）;各等位基因在两组间比较差异具有显著性意义（P〈0.05）;A等位基因与PE呈负相关（RR=0.475,P〈0.05）;G等位基因与PE呈正相关（RR=5.818,P〈0.05）。结论α-纤维蛋白原基因Thr312Ala位点G等位基因与肺动脉栓塞有正关联,肺动脉栓塞死亡者中GA、GG基因型分布频率较高,本文数据可为相关研究提供参考。