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.     

Plasminogen (PLG) polymorphism in northern Japanese: confirmation of PLG*M6 allele

Plasminogen (PLG) phenotyping has been performed on 450 unrelated individuals from northern Japan, using wide-scale ultrathin layer polyacrylamide gel isoelectric focusing combined with immunoblotting. One common phenotype and six rare ones were observed. The rare phenotypes included the recently detected allele PLG*M6 in a new combination with PLG*M5 allele. The estimated allele frequencies for PLG*A, PLG*A3, PLG*M2, PLG*M5, PLG*M6, PLG*B, and PLG*B2 were 0.961, 0.009, 0.001, 0.016, 0.001, 0.003, and 0.009, respectively.     

Jordanian population data on the PCR-based loci: LDLR, GYPA, HBGG, D7S8 and GC.

Genotype and allele frequency distributions for PM polymerase chain reaction (PCR)-based genetic markers were determined in a Jordanian sample population. Results were obtained using the AmpliType PM PCR Amplification and typing kit. All loci were in agreement with the Hardy-Weinberg equilibrium expectations. The predominant alleles for LDLR, GYPA, HBGG, D7S8 and GC loci were B, A, B, A and C respectively. No statistically significant variation was detected in allele frequencies of these loci in Jordanians compared to that in Israeli Arab, U.S Caucasian and Japanese populations. Data presented here can be used to estimate the frequency of a specific DNA profile in the Jordanian population for forensic analyses and paternity testing.     

C6-polymorphism of the sixth component of complement: application to paternity cases (author's transl)]

The results of a study of the polymorphism of the sixth component of human complement by means of isoelectric focusing in polyacrylamide gels with subsequent C-dependent lysis in an agarose overlay containing C6 deficient rabbit serum are reported. The allele frequencies obtained (C6A = 0.613, C6B = 0.379, C6R = 0.008) are in good agreement with those previously published. The mode of inheritance in 47 families with 173 offspring as well as 26 mother-child combinations is in agreement with a formal genetical model: "C6A, C6B, C6A1 and C6B1 at an autosomal locus". The inclusion of this system into a blood group expertise in Germany can be recommended.     

Properdin factor B-polymorphism. An indication for the existence of a Bf O-allele.

The polymorphism of the properdin factor B (Bf, C3-proactivator, GBG = glycin-rich-beta-glycoprotein) has been investigated by high voltage agarose gel immunofixation electrophoresis in 1115 unrelated persons from Southern Germany. Seven phenotypes were observed; the allele frequencies were calculated as BfS = 0.8094, BfF = 0.1790, BfSI = 0.0094, BfFI = 0.0022. A study of 94 parents with 98 children and 420 mother-child combinations showed no deviation from the assumed autosomal codominant mode of inheritance. In one additional family the findings suggested the existence of a silent allele at the Bf-locus.     

Automated estimation of the number of contributors in autosomal STR profiles

Estimating the number of contributors to mixed STR profiles can be complex. This study describes the nC-tool to assist DNA expert in this process. The nC-tool is based on the total allele count for PowerPlex® Fusion 6C profiles and showed improved performance when compared to the maximum allele count approach.     

Alleles responsible for ABO phenotype-genotype discrepancy and alleles in individuals with a weak expression of A or B antigens

ABO types obtained from evidentiary samples have been used effectively to obtain the initial information leading to the apprehension of culprits in Japanese criminal investigations. A simple ABO genotyping method using multiplex sequence-specific PCR and capillary electrophoresis was developed as a supplement to serological ABO typing. Limitations in predicting a phenotype based on genotype were evaluated using 1134 randomly selected Japanese peripheral blood samples. A concordance rate of 99.82% (1132/1134 samples) was found between genotypes and phenotypes defined as Groups A, B, AB, and O. Sequencing analysis revealed that one discrepant sample contained an O allele having a previously unreported point mutation at the primer binding site in exon 6, and another discrepant sample contained an O allele lacking the guanine deletion at nt 261 (the O301 allele). Therefore, the existence of such alleles must be given some consideration when predicting phenotype based on genotype.     

The BF F subtypes are detectable in the Ba fragment of factor B

The unanimous recognition of the two subtypes FA and FB of the BF*F allele has repeatedly been challenged. In the present investigation we are reporting about the unequivocal and simple detection of the subtypes on the Ba fragment of factor B by immunofixation isoelectric focusing after conversion with inulin. The common BF phenotypes F, S, and FS could be diagnosed in addition to the subtypes of BF*F which were observed in two regions acidic of the F major band. By comparison of standard phenotypes the subtypes in the Ba fragment corresponded to those of native factor B. All BF bands could be attributed to the Ba fragment by developing Western Blots with monoclonal antibodies directed against Ba. The distribution of the major BF phenotypes and alleles and the BF F subtypes in a population sample of 527 unrelated individuals from F.R.G. was in Hardy-Weinberg equilibrium. The allele frequency was determined to be 0.0731 for BF*FA, and 0.1053 for BF*FB. The advantages of determining the subtypes on the Ba fragment are: broadening of the FA/FB corridor, a more reliable diagnosis of phenotypes, improved distinction between homozygous FA and heterozygous FAFB types, and recognition of common BF phenotypes as well as subtypes in aged sera. It is suggested that the problem in the designation of BF F subtypes by different groups should be resolved by an international reference typing.     

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.     

Determination of the effects of alcohol dehydrogenase (ADH) 1B and ADH1C polymorphisms on alcohol dependence in Turkey

Alcoholism is a complex genetically influenced disorder which refers to alcohol abuse and alcohol dependence. There are controversial results on the role of gene polymorphisms in alcohol dependence in the literature. Differences in population groups and selective inclusion criteria for alcohol dependence may affect results. In this study, we investigated the role of ADH1B Arg48His (rs1229984) and, ADH1C Ile350Val (rs698) gene polymorphisms in Turkish population. 100 healthy volunteers and 75 patients who were admitted to Ege University Alcohol Dependence Unit enrolled in the study. We found significant increase both in ADH1B (Arg48His) polymorphism Arg allele and Arg/Arg genotype frequency in patients. No profound connection between alcohol dependence and ADH1C Ile350Val gene polymorphism was detected. Alcohol dependence is an important health problem that depends on many genetic and environmental factors but we think that it is possible to interpret genetic risk for developing early diagnostic methods and treatment strategies by comprehensive linkage and association studies.     

Characterization of the variant allele 9.2 of Penta D locus

DNA profiles of forensic cases of Córdoba Province, Argentina, typed by PowerPlex 16 kit (Promega), have shown in the Penta D locus few samples with a variant allele migrating as an off ladder between alleles 9 and 10. In order to determine the molecular basis of the new variant allele, three samples were subject to polymerase chain reaction amplification of the Penta D locus by monoplex, and were further purified and sequenced. The sequence analysis revealed that the off ladder allele has ten repeats motifs AAAGA as allele 10, with three nucleotides (TAA) deletion in the 3' flanking region, 128 nucleotides after the last repeat. Therefore, the variant allele could be explained by a deletion of allele 10, and was designated 9.2. Mse I digestion assay allows to corroborate allele 9.2 without sequencing. A population study in Córdoba Province indicates that allele 9.2 of Penta D locus has a frequency of 0.0063.     

A new allele of the short tandem repeat (STR) locus, CSF1PO.

CSF1PO is one of the thirteen core loci used for the CODIS database, and alleles reported for this short tandem repeat (STR) locus contain from 6 to 15 repeats of the tetranucleotide AGAT. Screening of DNA from 76 individuals by gel electrophoresis and silver stain detection yielded one sample that contained a rare, off-ladder CSF1PO allele; an allele larger than CSF1PO15 was detected in a heterozygote that also contained a CSF1PO10 allele. Capillary electrophoresis analysis using GeneScan software demonstrated that the variant allele contained four bases more than CSF1PO15. Following agarose gel electrophoresis to separate the two alleles of the heterozygote and cycle sequencing using dye terminators, sequence analysis showed that the variant, which was otherwise identical to the CSF1PO GenBank sequence, contained exactly 16 AGAT repeats. These results demonstrate the existence of an additional CSF1PO allele, a previously unreported size variant, CSF1PO16.     

人类补体第八成份B基因TapI限制性片段长度多态性研究

采用补体第八成份B基因(C8B)11号内含子特异性PCR扩增技术和直接TaqI消化处理,检测了121份中国成都地区汉族群体的DNA样本的C8B TaqI限制性片段长度多态性,首次证实了C8B Taq RFLP在中国群体的分布,其基因频率分别为C8B TaqI 2.6*=0.4463和C8B Taq 1.8,0.8*=0.5537.经计算其表现型频率分布符合Hardy-Weinberg定律,并与德国群体在该基因座的分布进行了比较.     

The application of an automated allele concordance analysis system (CompareCalls) to ensure the accuracy of single-source STR DNA profiles

A powerful method for validating a scientific result is to confirm specific results utilizing independent methodologies and processing pathways. Thus, we have designed, developed and validated an automated allele concordance analysis system (CompareCalls, patent pending) that performs comparisons between two independent DNA analysis platforms to ensure the highest accuracy for allele calls. Application of this system in a quality assurance role has shown the potential to eliminate greater than 90% of the STR analysis required of a DNA data analyst. While this system is broadly applicable for use with any two independent STR analysis programs, either prior to or following human data review, we are presenting its application to data generated with the ABI Prism Genotyper software system versus data generated with the SurelockID system. With the automated allele concordance analysis system, the GeneScan DNA fragment data generated from an ABI 377 gel image are analyzed in two independent pathways. In one analysis pathway, the GeneScan data are imported into Genotyper software where STR labels are assigned to the fragment data based upon the criteria of the Kazam 20% macro. The "Kazam" macro provided with the Genotyper program works by labeling all peaks in a category (or locus) and then filtering (or removing) the labels from peaks, such as those in stutter positions, that meet predefined criteria. In the second pathway, the GeneScan data are imported into the SurelockID analysis platform where STR labels and error messages are assigned to the fragment data based upon hard-coded allele calling criteria and quality parameters. The resulting STR allele calls for each analysis platform are then compared, utilizing the automated allele concordance analysis system. Any differences in the STR allele calls between the two systems are flagged in a discordance report for further review by a qualified DNA data analyst. The automated allele concordance analysis system guides the DNA data analyst to the discordant data generated by either analysis platform. Additionally, the analyst is also directed to data that are of less than pristine quality which may have an increased potential for errors in interpretation by either analysis platform or by a human DNA data analyst. Implementation of an automated allele concordance analysis system will yield high-quality data for CODIS and free the human DNA data analyst to perform other critical duties within the laboratory.     

Population genetic data of eight tetrameric short tandem repeats (STRs) in Casablanca resident population to use in forensic casework

The allele frequencies for eight short tandem repeat (STR) loci HUMvWA, HUMFES/FPS, HUMF13A, HUMF13B, HUMTHO1, HUMTPOX, HUMCSF1P0, HUMLPL included in Geneprint STR kits were obtained from 234 unrelated individuals in Casablanca.     

突变情形下亲权指数计算的新方法

目的介绍一种常染色体共显性遗传标记突变时的亲权指数（paternity index,PI）计算法。方法假定突变的等位基因在传给孩子之前以一突变概率发生突变．然后发生分离以1／2的机会传给孩子。而随机男子提供某一等位基因给孩子的概率为等位基因频率。一个案例中考虑仅有一次突变,通过比较亲代和子代的等位基因确定突变等位基因,分别算出被检男子和随机男子是孩子生父时,出生该孩子的基因型的概率,并计算出相应的PI值。结果推导出三联体、二联体和孩子失踪案中有突变（包括母亲突变）时PI的计算公式。结论本方法中考虑突变时的PI计算方法易于理解和掌握,在亲权鉴定实践中易于使用。     

中国汉族群体人类补体C8A多态性

采用免疫沉淀、SDS-聚丙烯酰胺凝胶电泳 (SDS- PAGE)、被动转印及酶免分析 ,研究了人类补体 C8A等位基因频率在成都地区汉族群体中的分布。 12 1份样本被分为 3种常见型 ,即 C8A- A、C8A- B及 C8A- AB,由两个等位共显性基因 C8A * A及 C8A* B控制 ;同时发现了 2个稀有亚型 ,即 A3亚型及新发现的 Ax亚型。等位基因频率为 C8A* A=0 .5 0 83,C8A* B=0 .4835 ,C8A*稀有型 =0 ,0 0 83。说明 C8A多态性在中国群体中具有良好的分布 ,个人识别率(DP)达到 6 1.14% ,可用于法医学个人识别及亲子鉴定     

Detection of the rare PGM1(3) allele. Further biochemical and genetic characterization of the PGM1(3) isozymes

A family possessing the rare PGM1(3) allele has been found in North Carolina, and criteria for the electrophoretic separation and accurate typing of the PGM1(3) isozymes are outlined. The PGM1(3) isozymes detected proved to be useful in helping to determine parentage in an incest investigation. The pattern of segregation of the PGM1(3) allele in four generations of this family and thermostability studies on the PGM1(3) isozymes are presented.     

Distribution of alleles of 12 STR loci in Tamil population (south India)

Distribution of allele frequencies for 12 STR loci (CSFIPO, TPOX, THO1, F13A01, FESFPS, vWA, D16S539, D7S820, D13S317, HPRTB, F13B and LPL) has been studied for the first time in unrelated Tamil (south India) population.     

DNA的apoB位点扩增片段长度多态性的检测及其在法医学中的应用

用PCR方法对100例无关个体血样的apoB位点扩增片段进行研究,已发现有11个等位基因,片段长度分布于600～1100bp之间,基因频率为0.005～0.525,杂合度为70％。家系分析及对人体血液、血斑、精液、精斑、混合斑、带毛囊毛发及其它各种有核细胞组织的研究表明,该技术在个人识别及亲子鉴定上均能发挥重要作用。