Sibling incest and formulation of paternity probability: case report

Paternity determination of a fetus whose mother was admitted to an institution for the welfare and health of handicapped persons was requested of us by a doctor and lawyer of the institution. The fetus was recovered by a legal artificial abortion based on the Act on Maternity Health and Welfare (Japan) with the permission of the custodian. Commercially available MCT118, HLADQA, PM, and 9 STRs were tested for DNA samples from the fetus, the mother, her younger brother, her father, her grandfather, and 4 staff members of the institution. Only the brother was not excluded and the paternity probability was estimated at 99.857% on the basis of newly formulated expressions for multiallelic loci on the assumption of sibling incest. We concluded then that the fetus was fathered by the brother. DNA fingerprinting with multilocus and single locus minisatellite probes which were performed to confirm the paternity also support the conclusion. Bandsharing frequencies between the family members, however, did not necessarily reflect their actual kinship, which findings suggest that multilocus DNA fingerprinting requires further accumulation of data for consanguineous cases such as incest. Universal formulation for calculating paternity probability for a sibling incest case on the basis of multiallelic monolocus polymorphisms is also presented.     

Application of single-locus hypervariable region DNA probes to deficiency cases in paternity testing

Summary Seven DNA probes which recognize single-locus hypervariable region (HVR) were applied to a paternity test in which the putative father and his wife were deceased. Three legitimate children, an illegitimate child and her mother were available for analysis. The cumulative paternity index of the illegitimate child derived from 15 conventional blood group markers was 18.71 and from 7 DNA probes 92,572.08, that is, 4,948 times higher than the former. Thus the DNA analyses gave nearly conclusive evidence that the putative father was the biological father of the child. The application of highly discriminating polymorphisms of DNA which recognize single HVR loci is considered to be extremely informative in cases of disputed parentage.     

Abstammungsbegutachtung mit der RFLP- und STR-Analyse

The combination of restriction fragment length polymorphism (RFLP) and short tandem repeat (STR) analyses for paternity analysis is presented. The two methods were compared by investigating 113 paternity cases. RFLP analysis was done using the single locus probes YNH24, MS31 and MS43A and for STR investigations the Identifiler Plus kit was employed. The lowest paternity probability obtained via RFLP analysis was 98.936% compared to 99.99844% when using STR analysis and the highest values were 99.9996 (RFLP) and >99.999999% (STR). Using 3 single locus DNA probes the paternity probability was <99.9% in 45.5% of the cases, while STR analysis always led to at least 99.9%. In 36 cases the father was excluded. Using STR analysis between 4 and 12 exclusions out of 15 investigated loci per case were observed. In 14 cases (39%) RFLP analysis alone did not yield the 3 exclusions necessary for exclusion of paternity. In summary it could be shown that in all cases both STR analysis alone and the combination of STR and RFLP investigations led to results which conformed to the requirements of the German guidelines.     

Pränatale Paternitätsbestimmung mittels PCR-VNTR-Polymorphismen

A case of prenatal paternity diagnosis where the woman had been raped is presented. PCR-VNTR polymorphisms were investigated on material which was obtained by biopsy during the 10th week of pregnancy. After DNA extraction nine PCR-VNTR polymorphisms were examined and the husband of the pregnant woman could not be excluded as the biological father of the child with a high degree of certainty. The ethical aspects associated with this problem will be discussed.     

DNA investigations on fetal material from paternity cases

Summary Three paternity cases have been investigated where DNA was extracted from fetuses (age: 8–10 weeks old) after interruption of pregnancy. In each case it was possible to clearly identify the putative father using 5 or 6 single locus probes (SLP's). Fetal bands (SLP) could be clearly identified from mixtures of placental and fetal DNA by comparison with the maternal and paternal bands. However, it was very difficult to resolve the fragment patterns of tissue mixtures with one multi locus probe (MLP), because of band overlap. Another advantage of using SLP's was that biostatistical calculations could be carried out and very informative Essen-Möller values for the probability of paternity were obtained.     

Evaluation of the paternity probability on an application of minisatellite variant repeat mapping using polymerase chain reaction (MVR-PCR) to paternity testing

Minisatellite variant repeat (MVR) mapping using polymerase chain reaction (PCR) was applied to a practical case of paternity testing to evaluate the paternity probability. In order to obtain single allele mapping by allele-specific MVR-PCR, three flanking polymorphic sites for each of the MS31A and MS32 loci were investigated and all three individuals were typed as heterozygous for at least one flanking polymorphic site at each locus. Allele-specific MVR-PCR was then performed using genomic DNA. It was confirmed that one allele in the child was identical to that from the mother and the other one in the child was identical to that from the alleged father. Mapped allele codes were also compared with those in the database by dot-matrix analysis, and no identical allele was found although some motifs were shared with Japanese alleles. The paternity index and the probability of paternity exclusion in the case at these two MVR loci were calculated using the presumed values of the allele frequencies. These studies seem to illustrate the practical value of MVR mapping of MS31A and MS32 loci in paternity testing.     

The Statutory Framework: A Checklist of Federal and State Drug Crimes and Defences to Them

A mother killed her two children and the woman that her husband preferred. Her actions and motives paralleled those of Medea, in the various versions of that myth. Her actions and intentions were interpreted in quite different ways by her husband, by the police, by each of the psychiatrists who examined her, by the jury, by the sentencing judge and ultimately by the Court of Appeal.     

产前亲子鉴定方法研究(附23例报告)

目的对23个产前案例进行亲子鉴定。方法超声监视下行羊膜穿刺术,抽取羊水30~40ml。离心收集羊水沉渣后提取其基因组DNA,同时抽取其父母双方外周血基因组DNA。应用毛细管电泳技术和五色荧光复合扩增的方法,检测所有DNA样本的16个STR基因座基因型。结果所有羊水基因组DNA均来自独立个体,无母体DNA的污染。三联体分析显示23个案例中17例为肯定亲权关系,亲子关系概率均大于0.9999,6例确定为排除亲权关系,平均排除(位点数)指标为7.67个。二联体分析显示23个案例中17例肯定父权的平均亲子关系概率为0.9997以上,6例排除亲权关系的平均排除(位点数)指标为5个,但其中1例的排除位点只有1个。结论16个STR位点的多重荧光扩增方法在对羊水中母体DNA的污染程度进行评估的同时,可以准确、可靠的应用于产前亲子鉴定。在检测单亲鉴定案例时,若排除(位点数)指标小于2时必须补充母亲样本或增加检测的STR位点指标数,直至得出明确结论。     

UT LIFESTAR-rising: to new heights for 20 years

While Christmas shopping with her husband in December, a 39-year-old east Tennessee woman began to feel faint and collapsed. Her husband took her to a local hospital, where she was reported to be in ventricular tachycardia. She was pale to cyanotic in color, diaphoretic, nauseated, and experiencing acute chest pain. Critically ill, the patient was experiencing an acute myocardial infarction (MI) and needed immediate intervention.     

Paternity evaluation in cases lacking a mother and nondetectable alleles

In parentage testing the formulae for computing paternity index and exclusion probability generally ignores the presence of nondetectable alleles at the loci tested. In contrast, it is now known that even when paternity testing is done with hypervariable DNA markers, nondetectable alleles should not be ignored. This work presents simple formulae needed with this consideration, to analyze paternity evaluation from DNA markers in cases where the mother of the disputed child is unavailable for testing. It is shown that even a modest frequency of nondetectable alleles (e.g., 2–5% per locus) may have a substantial impact on the paternity index when the child and/or the alleged father exhibits a single-banded DNA profile at a locus. Use of such formulae can generate a high probability of exclusion and a high paternity index when multiple independently segregating hypervariable DNA markers are used.     

DNA-Untersuchung von Hautepithelzellen asserviert am Rollkragenpullover und am Hals des Opfers

A 34-year-old woman who was separated from her husband, was killed by him by strangulation and several stab wounds in the chest. The post-mortem examination was carried out approximately 5 h after death. At the place of discovery one swab of her right and one of her left neck side were taken. Additionally, a swab was taken out from the front side of the collar of her polo-neck pullover. It was obvious that the skin of the neck below the larynx showed a fabric pattern leading to the assumption that the collar was temporarily pressed between the hands of the suspect and the neck of the victim. DNA typing (short tandem repeat-PCR) of these swabs led to the detection of mixed patterns for the swabs of the neck and the collar which matched the patterns of the victim and the suspect.     

Fetal male lineage determination by analysis of Y-chromosome STR haplotype in maternal plasma

The aim of this study is to determine the fetus Y-STR haplotype in maternal plasma during pregnancy and estimate, non-invasively, if the alleged father and fetus belong to the same male lineage. The study enrolled couples with singleton pregnancies and known paternity. All participants signed informed consent and the local ethics committee approved the study. Peripheral blood was collected in EDTA tubes (mother) and in FTA paper (father). Maternal plasma DNA was extracted by using NucliSens EasyMAG. Fetal gender was determined by qPCR targeting DYS-14 in maternal plasma and it was also confirmed after the delivery. From all included volunteers, the first consecutive 20 mothers bearing male fetuses and 10 mothers bearing female fetuses were selected for the Y-STR analysis. The median gestational age was 12 weeks (range 12–36). All DNA samples were subjected to PCR amplification by PowerPlex Y23, ampFLSTR Yfiler, and two in-house multiplexes, which together accounts for 27 different Y-STR. The PCR products were detected with 3500 Genetic Analyzer and they were analyzed using GeneMapper-IDX. Fetuses’ haplotypes (Yfiler format) were compared to other 5328 Brazilian haplotypes available on Y-chromosome haplotypes reference database (YHRD). As a result, between 22 and 27 loci were successfully amplified from maternal plasma in all 20 cases of male fetuses. None of the women bearing female fetuses had a falsely amplified Y-STR haplotype. The haplotype detected in maternal plasma completely matched the alleged father haplotype in 16 out of the 20 cases. Four cases showed single mismatches and they did not configure exclusions; 1 case showed a mutation in the DYS 458 locus due to the loss of one repeat unit and 3 cases showed one DYS 385I/II locus dropout. All mismatches were confirmed after the delivery. Seventeen fetuses’ haplotypes were not found in YHRD and one of them had a mutation, which corresponded to the paternity probability of 99.9812% and 95.7028%, respectively. Three fetuses’ haplotypes occurred twice in YHRD, which corresponded to paternity probability of 99.9437%. In conclusion, high discriminatory fetal Y-STR haplotype could be determined from maternal plasma during pregnancy starting at 12 weeks of gestation. All male fetuses could be attributed to the alleged father male lineage early in pregnancy. The high probability of paternity associated with each case suggests that the relationship is not random and this strategy can be use as an alternative for male fetal kinship analysis.     

A silent allele in the locus D19S433 contained within the AmpFℓSTR® Identifiler™ PCR Amplification Kit

We present two cases where a single locus mismatch was found in the locus D19S433 using the AmpF?STR® Identifiler? PCR Amplification Kit (Applied Biosystems) (Identifiler Kit) during paternity and maternity tests. This mismatch differed from the mismatch pattern where there is usually a one repeat difference. We designed forward and reverse primers so that they were positioned further away from the primer set contained in the Identifiler Kit. The results showed the existence of a silent allele 13 in both families, due to a point mutation that changed guanine to adenine at 32 nucleotides downstream from the 3′ end of the AAGG repeat sequences in all four members. A single locus mismatch due to a silent allele may occur in any locus using any kit. Accordingly, we should pay attention to this silent allele when carrying out human identification and parentage analysis.     

Non-pathological complete paternal uniparental isodisomy of chromosome 2 revealed in a maternity testing case

International Journal of Legal Medicine - We present a duo paternity test case to assess the biological relationship between a woman and her female child. After analyzing 57 autosomal and 19...     

Homicidal strangulation by victim’s own artificial hair extensions

A 24-year-old woman was strangulated by her husband using his wife’s own 60-cm long artificial head hair integrations. During a domestic conflict, she was strangulated while lying in bed in a prone position. Her husband wrapped her hair around her neck and pulled it with one hand while fixing her head with the other hand. Due to the soft and broad nature of the ligature, the neck skin presented scarcely any ligature marks and showed a horizontal pale area in the middle of the neck above which the neck and face were congested; the skin showed extensive petechial haemorrhages. The autopsy revealed no internal injuries except two small haematomas in the soft tissue on each side of the neck. These were located under the pale area of the skin. The larynx and hyoid bone were intact. No natural disease was found. The toxicological analyses showed negative results. This is the first report of a homicidal strangulation by means of the victim’s own artificial hair extensions.     

Assisted suicide bordering on active euthanasia

A 44-year-old woman was almost completely paralysed after a severe brainstem haemorrhage. Even after several years of efforts at rehabilitation, she remained completely dependent on the help of others. However, a special device enabled her to administer (after careful preparation) liquids through the PEG catheter despite her poorly coordinated movements. Four years after the stroke, the woman joined a right-to-die society with the wish to bring her life to an end. A doctor working with this organisation prescribed her a lethal dose of pentobarbital. In the presence of her husband and her companion from the organisation, the woman administered herself the lethal substance by means of the device. On the basis of the fact that she herself had switched the device on this death was classed as (assisted) suicide.     

Forensic evaluation of HUMCD4: An Italian database

The YTTTC pentanucleotide short tandem repeat polymorphism HumCD4 was studied in an Italian population sample. PCR products were compared to an allelic ladder by manual PAGE and silver staining. A total of 6 alleles ranging from 5 to 12 repeats were represented in the analysed sample, of which 3 alleles (10, 6 and 5 repeats) were predominant and displayed a combined frequency of 0.91. Successful amplification was obtained from different sources such as blood and urine stains, teeth and paraffin embedded tissues. Results were also determined in cases of severely degraded DNA. We consider that the HUMCD4 polymorphism may be a useful tool for individual identification, paternity testing, population studies and have also employed this locus to monitor engraftment of bone marrow transplantation.     

Frequency data on four tetrameric STR loci D18S1270, D14S608, D16S3253 and D21S1437 in a Korean population

We present the results of a population study in Korea for four new tetrameric short tandem repeat (STR) loci employing multiplex PCR amplification, polyacrylamide gel electrophoresis of the PCR products and silver staining, which allow single base pair resolution and rapid typing. The loci tested were D18S1270, D14S608, D16S3253 and D21S1437 and all loci showed no significant deviations from Hardy-Weinberg equilibrium in more than 100 unrelated Koreans. This allelic frequency data can be used in forensic analyses and paternity tests to estimate the frequency of a multiplex PCR based DNA profile in the Korean population. Received: 13 January 1999 / Received in revised form: 14 June 1999     

Non-invasive prenatal paternity testing from maternal blood

Prenatal paternity analysis can be performed only after invasive sampling of chorionic villi or amnionic fluid. Aiming to enable noninvasive paternity testing, we attempted to amplify fetal alleles from maternal plasma. Cell-free DNA was isolated from plasma of 20 pregnant women and amplified with ampFLSTR Identifiler and ampFLSTR Yfiler kits. Unfortunately, autosomal fetal alleles were heavily suppressed by maternal DNA, and the only locus that was reliably amplified with AmpFLSTR Identifiler kit was amelogenin, which revealed only fetal gender. Much better success was obtained with AmpFLSTR Yfiler kit, which, in the case of male fetuses, successfully amplified between six and 16 fetal loci. All amplified fetal alleles matched the alleles of their putative fathers, confirming the tested paternity. To the best of our knowledge, this is a first report of noninvasive prenatal paternity testing. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.