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Paternity testing by PCR-based STR analysis.

We present application of polymerase chain reaction (PCR)-based short tandem repeat (STR) system for use in paternity testing. The process involves a single tube multiplex PCR of 9 STR loci on different chromosomes, in conjunction with Amelogenin sex test and internal size standards, followed by using an automated DNA sequencer to detect amplified products. The results showed that this system provided unambiguously reliable results. In addition, the method is useful for routine use in that it is robust and reproducible and provides a reliable means of paternity testing.

Humans↗

Automated fluorescent detection of a 10 loci multiplex for paternity testing.

The discovery of new highly efficient tetra repeat STR loci, development of fluorescence multicolour dye technology and capillary electrophoresis have made it possible to amplify ten loci in a single reaction. This combination provides an extraordinary effectiveness of simultaneous amplification and detection. With this method it became possible to determine individual identity and paternity at an enhanced level of precision and accuracy in 1 to 2 days with a high biostatistical probability. This review demonstrates the role of automated fluorescent multicolour dye genotyping technology in forensic paternity testing.

Electrophoresis, Capillary↗

Possibility of paternity testing using RFLP analysis on a very small amount of material.

Paternity testing by DNA analysis was carried out using dental pulpal and chorionic villous tissue from two children respectively, and fresh blood samples obtained from the alleged parents. DNA was extracted spectroscopically from the pulp of an upper wisdom tooth (16 micrograms) and the chorionic villi (53 micrograms). The RFLP method was used for DNA analysis of the parent-child relationships because both of the DNAs extracted had a high molecular weight. Distinct bands were detected with 32P-labelled multi-locus (Myo) and single locus (pYNH24) DNA probes. In the case of the dental specimen all of the bands of the child's DNA were found to be derived from either of the alleged parents, demonstrating a consistent parent-child relationship (the probability of established paternity was 99.86%) whilst in the case of the villous specimen the father-child relationship was denied. This procedure can provide much information using very little material for analysis but where the samples are in a good condition.

Child↗

The physician as fortune teller: a commentary on "The ethical justification for minimal paternalism.

...I shall summarize my argument to this point. 1) A new technology -- medicalizing life choices -- introduces additional sources of uncertainty into the physician-patient relationship. The implications for the authority-autonomy tension of that relationship are not fully worked out, as is evidenced by differing views on professional obligations. 2) The warrants for paternalism are complex, ranging from a pragmatic response in the face of uncertainty to a proactive, highly directive determination of the patient's best interests. 3) Although some proponents of autonomy argue that it entails positive rights to intervention and information, the "strong sense" of autonomy is not the prevailing view. 4) The as yet unclear benefits of knowledge about one's genetic makeup are even less clear in the case of HD. Strong circumstantial evidence of risks of harm from disclosure of unfavorable test results have not been dispelled by early experience with use of the HD test. 5) A cautious approach in the use of HD test seems warranted, even at the cost of restricting autonomy. Legitimating that approach by reference to traditional ethical theory is a necessary element in the transfer of the new technology to the wider clinical setting. 6) While PMP [the principle of minimal paternalism] attempts to offer such an approach, some questions do not appear to have been resolved or fully addressed by this formulation.

Altruism↗

Patient care and paternalism: dilemmas of family practice.

From the clinical records of a country doctor, this vignette concerns a teenaged girl who, having refused treatment, is persuaded, under near duress, to accept a regimen that her family physician considers best for her. Although apparently arrogant paternalism, the practitioner's approach proves, on reflection, to possess considerable merit. The author discusses the ethical principles that have led to rejection of paternalism in the West. Formulated as absolute maxims, they soon require, like all absolutes, a multitude of explanations and additions. Some logical, social, and other "exceptions" are briefly mentioned, because the old doctor's intuitive actions seem to have oddly coincided with a number of them. Yet the questions remain: Should this medical practitioner have become so deeply involved? Should he have interfered with his patient's autonomy to the extent he did? Was he justified?

Adolescent↗

[Blood group expertise in the clarification of disputed paternity].

The article gives a review of the legal situation of the illegitimate child in the GFR and of the principles in the statement or refutation of paternity. The directives, valid for the blood group expertise on parentage and the biostatistical evaluation of blood group results are discussed. Finally a survey of the value of a positive or negative biostatistical hint for paternity is given.

Blood Group Antigens↗

[Cantonese population data of the tetranucleotide repeat locus D7S809 and its application in paternity test].

The tetranucleotide repeat locus D7S809 was investigated in Cantonese population by polymerase chain reaction (PCR) amplification, subsequent polyacryramide gel electrophoresis and silver staining. 14 alleles and 50 genotypes were detected in 190 sample individuals. All alleles differed in size by 4 bp repeat. No interalleles were found. The heterozygosity, power of discrimination and chance of non-paternal exclusion were 0.8613, 0.9645 and 0.7184 respectively. No significant deviations from Hardy-Weinberg equilibrium were found. The D7S809 was a highly polymorphic, stable and accurate type locus and had been successfully applied to 100 paternity test cases.

Alleles↗

[Evaluation of 9 STR loci in paternity identification].

9 STR loci obtained by four-dye fluorescent labeling technique in paternity identification provides much information at one test and the cumulative chance of exclusion gets up to 0.9999. Our result of 268 paternity test cases shows that there are at least two incompatible loci in all Mother-Child-Alleged Father (M-C-AF) exclusive cases. To those unexclusive cases, The RCP all reaches international standard. It is suggested that more STR loci be used for accurate test in Child-Alleged Father(C-AF) case.

China↗

Paternity testing in case of brother-sister incest.

We performed a paternity test in a case of incest between brother and sister. DNA from blood samples of the alleged parents and their two children was obtained with Chelex DNA extraction method and quantified with Applied Biosystems QuantiBlot quantitation kit. Polymerase chain reaction (PCR) amplification of DNA samples was performed with AmpFlSTR SGM Plus PCR amplification kit and GenePrint PowerPlex PCR amplification kit. The amplified products were separated and detected by using the Perkin Elmer's ABI PRISM trade mark 310 Genetic Analyser. DNA and data analysis of 17 loci and Amelogenin confirmed the suspicion of brother-sister incest. Since both children had inherited all of the obligate alleles from the alleged father, we could confirm with certainty of 99.999999% that the oldest brother in the family was the biological father of both children. Calculated data showed that even in a case of brother-sister incest, paternity could be proved by the analysis of Amelogenin and 17 DNA loci.

Amelogenin↗

Utilization of cytogenetic technics in paternity investigation.

The joint analysis of several genetic markers in cases of paternity investigation renders possible a cumulative probability of 99.7% of chance of exclusion of a falsely accused father. The role of heteromorphisms of the Y chromosome size was appraised in this work, with more than one genetic marker, in 20 expertise examinations in paternity investigation, where the children were male. The results found with this method, in association with the research on erythrocytic and leucocytic antigens showed the exclusion of two falsely accused men. Cytogenetics analysis with Giemsa stain in combination with leukocyte (HLA system) and erythrocyte antigens investigation demonstrated the exclusion of two men falsely accused.

Cytogenetics↗

Quantitative indices in paternity cases.

The study discusses the basic quantitative indices used as a standard method in foreign professional literature dealing with paternity cases. They are as follows: 1. mean probability of exclusion (PE) which characterizes the informative value of the experts opinions and is the same in all the disputes evaluated by this expert. 2. relative frequency of men chosen at random from the population and excluded at given phenotype of mother and child (RME). 3. probability of paternity (PP) for particular trio: mother-child-the accused man. Hereinafter the results of our studies in the HLA laboratory in Olomouc from 1976-1991 are introduced.

Blood Group Antigens↗

[Investigation of algorithm for the calculation of probability of paternity likelihood using personal computer program, including the application to parentage testing in the decreased party].

Algorithm for the computerized calculation of probability of paternity likelihood was investigated. The probability is calculated by Essen-Möller's formula as W = X/(X+Y) = 1/(1 + Y/X). The X value is also given as X = Hl, m, n/Kl, m, where kl, m and Hl, m, n are the probabilities of mother-child and mother-child-father combinations, respectively. In this study, four functions as F(PQ) = [1-(P not equal to Q)] x p x q, Z(RS) = (1- (R = S)), K(PQ,RS) = 1/2([(R = P) + (R = Q)].s + [(S = P) + (S = Q)].r).F (PQ)/Z(RS) and H(PQ, RS, TU) = 1/4([(R = P) + (R = Q)] [(S = T) + (S = U)] + [(S = P) + (S = Q)] [(R = T) + (R = U)]) x F(PQ).F(TU)/Z(RS) were created, where PQ, RS and TU were the genotypes of mother, child and the alleged father, P, Q, R, S, T and U were their alleles, and p, q, r, s, t and u were the allele frequencies. The equality or inequality in parenthesis was the relation operator which gave -1 or 0 when the expression was true of false, respectively. Then, three formulae as Y = n sigma k = l F([TU]k), Kl, m = 1 sigma i = l m sigma j = l K ([PQ]i, [RS]j) and Hl, m, n = l sigma i = l m sigma j = l n sigma k = l H([PQ]i, [RS]j, [TU]k) were obtained, where [PQ]i, [RS]j and [TU]k were one of the mother's, one of the child's and one of the putative father's genotypes considered from their phenotypes, respectively. Using these formulae, the probability of paternity likelihood could be calculated in every case. These formulae were programmed in BASIC language using a personal computer. Algorithm for the calculation of the probability in the deceased party was also investigated.

Algorithms↗

Validation and uses of a Y-chromosome STR 10-plex for forensic and paternity laboratories.

Y-chromosome short tandem repeats (Y-STRs) provide valuable information in cases of rape and questioned paternity, and they allow for the genetic identification of male lineages. The present study validated a Y-STR 10-plex on the ABI PRISM 3100 Genetic Analyzer for use in forensic and paternity laboratories at Orchid Cellmark. Following optimization of the polymerase chain reaction, father-son pairs were analyzed to ensure that each pair generated identical haplotypes. This study demonstrated that the 10-plex is sensitive to 0.125 ng of input DNA and that female samples mixed with male samples did not interfere with Y-STR haplotyping. In a sample of 525 males, there were three instances of locus multiplication, two at DYS19 and one at DYS435. Overall, haplotype diversity was 0.996, suggesting that the 10-plex can effectively distinguish among male lineages.

Chromosomes, Human, Y↗

[Evolution of Supreme Court settlements relating to DNA evidence in paternity investigations].

The evolution of Supreme Court settlements relating to DNA evidence in paternity investigations was presented in this paper. It was found that its opinions have essentially changed during the last years as follows: 1) superiority of DNA evidence over other proof was perceived by the possibility of positive paternity confirmation, 2) attention has been turned to advisability of statistical analysis and its possible positive influence on emotional ties between parents and child, 3) opinion has been expressed about crossing of borders of free judicial estimations in cases of questioning DNA expertise, 4) attention has been turned to the possibility of utilisation of other tissues than blood or indirect genotyping.

Child↗

[Paternity testing and civil affairs].

Paternity testing is based on the genetic analysis of human polymorphism markers. Recently, studies of short tandem repeats (STR) after DNA amplification have led to increased standardisation and performance of the tests and have become the reference method. Thus, the results expressed in terms of probability achieve a high level of certitude, for example the probability of paternity can exceed 99.999%. All these analyses, which are performed according to a specific quality assurance program, are a very useful tool for courts of justice.

DNA↗

Determination of paternity disputes in the Niger delta region of Nigeria.

Results of paternity test, among disputants, carried out in our hospital between May 1992 and May 2002 were analysed. The cases were referred from Social Welfare Office, the Court and the Police. A total of twenty-four (24) cases consisting of 22 (91.7% ) groups of three each, including the disputed child, the mother, and the alleged father; and 2 (8.3% ) groups of four each, with one group consisting of the disputed child, the mother and two alleged fathers, while the other consisting of the mother, the alleged father and the disputed female twins. All the disputed victims were children (10 males and 14 females) in the age group 6 months to 8 years. The total cumulative paternal exclusion rate with our routine techniques was only 16.7% , all with the ABO blood group system. This is a far cry from the exclusion rates of about 97% for HLA testing, and 99.9% for DNA analysis. The available screening package in our hospital is therefore inadequate, and we advocate for the provision of more sophisticated and reliable techniques in designated centres across the country. This will go a long way in resolving the medicolegal and social problems encountered by the disputants and the victims.

Blood Group Antigens↗