Paternity testing by the detection of D1S80 VNTR using fluorescence image analyzer (Dualcolour system)

An improved method for DNA polymorphism typing of D1S80 VNTR locus and its application to paternity testing are described. For accurate estimation of the length of polymorphic DNA fragments, the size marker was labeled with fluorescence different from that of PCR primers, and co-electrophoresed as an internal standard. The dualcolour system of fluorescence image analyzer was used to detect the fragments and determine their size. This internal marker method could successfully overcome the problems of band pattern distortion and tailing, besides it allows easy and accurate interpretation of the DNA types. Our results indicate that the internal marker method is much more accurate than the method of using size marker in gel, even with the presence of distortion or tailing of the band patterns. Family studies applying this method showed complete agreement between the observed and predicated types.     

葡萄胎的亲缘关系鉴定

根据组织病理学特点及基因来源，葡萄胎可分为部分性葡萄胎和完全性葡萄胎。葡萄胎的类型及组织来源的判断除可采用组织病理学的方法外，还可采用STR基因座检测的方法。根据葡萄胎的类型、组织来源及受精方式，利用群体遗传学的原理和方法对葡萄胎进行亲缘关系鉴定。     

Paternity testing in a kidnapping case

A case of kidnapping involving the concept of paternity testing is described. The couple arrested and charged with kidnapping stated the child in their possession was their biological daughter. Serological tests were performed on liquid blood samples in an effort to determine if the couple could have been the biological parents of the child. Two genetic inconsistencies were discovered in the 15 blood marker systems analyzed.     

Paternity testing: blood group systems and DNA analysis by variable number of tandem repeat markers

Two recent paternity cases are reported. In the first case of paternity exclusion, deoxyribonucleic acid (DNA) restriction fragment length polymorphisms (RFLPs) on variable number of tandem repeat (VNTR) loci with multiple alleles were informative, as well as established systems of red blood antigens, red cell enzymes, serum proteins, and human leukocyte antigens. In the second case, in which both the alleged father and the first wife were deceased, the paternal genotype was determined by using genetic markers from the second wife and four children, which then were compared with the paternal alleles of the child in question, the plaintiff in this case. The high probability of paternity (0.999,998,7) made us conclude that the man probably was the actual father. The DNA analysis by VNTR probes appears to be quite valuable in the study of paternity cases.     

Population data on the forensic genetic markers: phosphoglucomutase-1, esterase D, erythrocyte acid phosphatase and glyoxylase I

Blood specimens from white and black sample populations from Baltimore, Maryland, were analyzed for the four most forensically important, polymorphic red cell enzyme systems-phosphoglucomutase-1, esterase D, erythrocyte acid phosphatase and glyoxalase I. The distributions of the phenotypes for each marker in each racial group were in Hardy-Weinberg equilibrium. The population data were similar to previously reported data for Whites and Blacks from different geographical locations within the United States.     

The esterase D polymorphism as revealed by isoelectric focusing in ultra-thin polyacrylamide gels

The polymorphism of human red cell esterase D (EsD) was studied using isoelectric focusing (pH 4-6) in ultra-thin polyacrylamide gels. Typing was possible without the EsD isozymes attaining true equilibrium focusing conditions. Using this single method, six phenotypes (EsD 1, 2-1, 2, 5-1, 5-2 and 5) could be recognized in the White population of south-east England. Family studies showed these to be controlled by three co-dominant alleles and the gene frequencies were calculated to be EsD1 0.8856; EsD2 0.0946 and EsD5 0.0198. For successful and reliable EsD typing by this method, the electrophoretic system must be carefully optimized with respect to the duration of electrophoresis and the temperature attained in the gel during the electrophoretic run.     

A case of paternity testing influenced by the silent allele of Rh erythrocyte groups

A paternity test is presented in which a father and his two children possessed an extremely rare amorphic gene R-29 (r,---). One of the children was determined to be illegitimate at the first trial as her Rh phenotype was R2R2(ccDEE) and the father's phenotype was R1R1(CCDee). At the Court of Appeal, however, the rare Rh gene r(---) was shown to be inherited from the father to the appellant child through extended tests including her brother whose phenotype was also R2R2(ccDEE). She was acknowledged to be legitimate.     

D18S51及其侧翼区3个SNP组成的SNP-STR在亲子鉴定中的应用

目的将一个单倍型区块内的遗传标记单核苷酸多态性(SNP)和短串联重复序列(STR)组成SNPSTR单倍型,调查其在成都汉族人群中的分布,并探讨其在特殊亲子鉴定案例中的应用价值。方法选取DNA联合索引系统(combined DNA index system,CODIS)中突变率较高的基因座D18S51,与其侧翼区的3个SNP位点(rs8089331、rs8094489、rs7236090)组成SNP-STR,通过巢式等位基因特异性PCR的方法获得SNP-STR单倍型,调查该单倍型在75名成都汉族人群中的分布,并应用于两例D18S51基因座不符合遗传规律的二联体亲子鉴定案件。结果成功建立SNP-STR分型方法,在成都汉族人群中共发现43种单倍型,多态性为0.948 6,并成功解决了两例二联体亲子鉴定案件。结论 SNP-STR具有良好的多态性,有望应用于特殊的亲缘关系鉴定。     

经耳郭软骨DNA检验认定亲缘关系1例

1案情 某年9月16日凌晨4时左右，某烟花厂发生爆炸．爆炸现场中心有南北两处炸坑。两炸坑坑沿间距离7．5m。南侧炸坑呈东西走向，长34．5m，宽30．5m，坑内有东西两处炸点，炸点坑深分别为3．9m和2．58m。北侧炸坑呈东南、西北走向，长32．3m，宽18．89m，坑内有两个炸点，炸点坑深分别为3．3m和3．42m。爆炸现场外发现尸体一具（尸体已被附近村民认领）。9月20日刘某报称其丈夫在该厂工作，现已失踪。     

Paternity determination when the alleged father's genotypes are unavailable.

In paternity testing using the DNA evidence, analysis of the deficiency case when the DNA profiles of the alleged father are not available is different from that of the case with complete evidence. In this paper, we describe how to evaluate and determine the paternity in the deficiency case, by comparing the paternity indexes of the true father and the falsely non-excluded man.     

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.     

Evidence for a "new" allele at the esterase D (E.C. 3.1.1.1.) locus

An apparently new EsD gene product (EsD*Düsseldorf) was detected by use of horizontal agarose gel electrophoresis (AGE), starch gel electrophoresis (SGE), and isoelectric focusing (IEF). The observed phenotype EsD (1-Düsseldorf) can be distinguished from any known EsD type.     

The use of isoelectric focusing (IEF) as a method for the combined phenotyping of erythrocyte acid phosphatase (EAP) and esterase D (EsD)

The simultaneous isoelectric focusing (IEF) in polyacrylamide gels (PAG) of erythrocyte acid phosphatase (EAP) and esterase D (EsD) allows the poor discriminating power (DP) of EsD to be usefully combined with a highly discriminating system EAP, such that a joint DP of 0.766 was achieved compared with PGM IEF DP 0.756. Focusing was carried out in a centrally flattened gradient containing ampholines (pH 4-6 and 6-8) and the chemical spacer 3-(N-morpholino) propanesulphinic acid (MOPS). It enabled the identification of six EsD phenotypes including the recently discovered EsD5 isozymes. The application of this method to casework bloodstains is discussed.     

A hybrid ampholyte focusing technique for esterase D subtyping of evidentiary material

An ultrathin-layer polyacrylamide gel isoelectric focusing technique that uses a composite of ampholytes from three commercial sources is described for subtyping esterase D. All common allelic products of esterase D were separated clearly. The technique described in this paper provides a higher conclusive call rate on known blood specimens (95.8%) and questioned bloodstains (69.7%) compared with continuous zone electrophoresis in agarose gels (89.9 and 37.6%, respectively).     

A forensic application of DNA typing. Paternity determination in a putrefied fetus.

Using minisatellite DNA probes that hybridize to a variable number of tandemly repeated loci, an individual-specific DNA fingerprint can be determined. In the case reported here, we succeeded in extracting high-molecular-weight DNA from a 3-month-old fetus discovered during the autopsy of a murdered 28-year-old pregnant woman reported missing 10 days earlier. The results of analysis of restriction-fragment-length polymorphisms showed that all bands present in the fetus's pattern, but absent in the mother's, matched only those of the putative father. Thus, the paternity of the victim's husband was ruled out.