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Family relationship proven by chromosomal and DNA examinations
International Congress Series 1239 (2003) S19 – S23
Family relationship proven by chromosomal and DNA examinations Gyo¨rgyi Bujdoso´ a,b,*, Pe´ter So´tonyi a,b, Andras Laszik a, Max P. Baur c a
Institute of Forensic Medicine, Semmelweis University, Budapest, Hungary b Research Unit Sponsored by Hungarian Academy of Science, Hungary c Institute of Medical Biometry and Epidemiology, University of Bonn, Bonn, Germany
Abstract As of today in Hungary, the live-birth rate is continuously declining, but the number of babies born from out of marriage relationship is increasing. According to the National Statistical Office, the later number represents 28% in 1999. In the past 30 years, we have been engaged in many cases where an adult wants to be informed about his or her ancestry. Out of these plentiful caseloads, we would like to present three interesting cases. Case 1: Identification of two stepbrothers with chromosomal and DNA investigations—without examining the parents. Case 2: Proved family relationship (father – son) with chromosomal translocation studies (22/Y). Case 3: After 55 years, family members were identified (blood groups, anthropology, chromosomes, DNA) of a prisoner of war (POW). D 2003 Elsevier B.V. All rights reserved. Keywords: Hereditary; Anthropology; Chromosome; DNA; Expert
1. Material and methods In our institution, we have access to perform blood group, anthropological, chromosomal and DNA examinations. In case of anthropological examination, the persons involved will be compared using characteristic features, with ample photo documentation [1,2]. Fourteen blood and serological groups were investigated. The chromosomes were cultivated from peripheral blood after 72 h. The applied staining methods were: Q and C
* Corresponding author. Institute of Forensic Medicine, Semmelweis University, Budapest, Hungary. 0531-5131/ D 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0531-5131(03)01053-7
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banding, rarely G banding if structural chromosomal abnormalities were detected [3– 8]. The DNA studies were done according to the recommendations of International Forensic Haemogenetic Society [9,10].
2. Case report 2.1. Case 1 V.M., a 31-year-old male, who was born out of wedlock, requested an examination on his genetic relationship to his so-called stepbrother. He had never known his father and he was well in his adulthood, when he found out about M.S., who was a 28-yearold man, born from a marriage. They speculated that they may have a common father. The hypothetical father has passed away, the mother did not give her consent to the examination; therefore, we could examine only the two assumed stepbrothers. During the chromosomal studies, we highlighted the chromosome 1, 9, 16 and Y with C banding and the chromosome Y with Q banding as well. The chromosomal studies showed V.M.’s assumption to be true, which was assured by DNA examinations (Table 1). 2.2. Case 2 In this case, an older man (whose name is the same as his son’s) wants to be sure that his offspring is really his son, because 50 years ago only fingerprint examination was possible, so he was in doubt in all his adult life about being the biological father. If this is proved, he can leave him a big share in his will. The mother of this son was not available, but it was not necessary, because the chromosome 22 was translocated to the chromosome Y in both tested persons. Besides that the paternity was proved absolutely, this case represents a rarity in the scientific literature. This case will be published in the near future with more minute details (Fig. 1). Table 1 DNA analysis PCR system
M.S.
V.M.
HUMTHO1 HUMvWF D18S51 FGA D8S1179 D3S1359 D21S11 SE33
6/6 14/16 12/15 22/22 15/15 5/14 29/30 17/26.2
6/9 14/18 16/19 19/21 13/15 5/14 29/33.2 26.2/28.2
DYS 19 DYS 390 DYS 392
16 24 11
16 24 11
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
Fig. 1.
Fig. 2.
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S22
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
Fig. 3.
Fig. 4.
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
S23
2.3. Case 3 In this third case, the presumed family members of a lost POW were found. Anthropological, blood group, chromosomal and DNA examinations were done on the POW (A.T.) as well as on his assumed brother and sister. A.T. become a prisoner in Russia during the WW II and has spent 55 years in a camp in a lunatic asylum. The persons investigated were: Andra´s Toma, suspected half-brother and sister, Ja´nos and Anna, from paternal side. You can see on these anthropological characters: the colour of the eyes, the upper eyelid’s hottentote wrinkles, the shape of the chin and its dimple at all three persons are the same. At Andra´s and Ja´nos, the shape of the eyebrows is the same. At Andra´s and Anna, the redness of the lower lip is the same. And now you can see their father (Fig. 2). At the chromosomal investigation, the size and the intensity of fluorescence at the Ychromosome were the same at both so we learned that they got a common father. Besides that, the choromosome numbers 4, 13, 14 and 21 were the same in all three of them (Figs. 3 and 4). The parallel DNA investigations proved and supported these findings. Biostatistical results PCR systems RFLP systems Y systems Blood serology In sum
99.98% 75.64% 97.08% 59.51% 99.9999%
References [1] G. Beitzke, H. Hosemann, P. Dahr, H. Schade, Vaterschaftsgutachten fu¨r die gerinchtliche Praxis, Verlag Otto´ Schwrtz et CO, Go¨ttingen, vol. II, 1978, pp. 58 – 59. [2] M. Bujdoso´ Gy, Anthropomorph-genetika Album, Kiege´szı´te´s az egyetemi jegyzethez, Rro Renovanda Cultura Hungeriae, Budapest, 1995. [3] T. Caspersson, L. Zech, C. Johannson, Quinacrine mustardfluorescence of human chromosomes, Exp. Cell Res. 61 (1970) 474 – 475. [4] P.P. Craig-Holmes, M. Shaw, Polymorphism of human constitutive heterochromatin, Science 1974 (1971) 702 – 704. [5] A.T. Summer, H.J. Evans, R. Buckland, New technique for distinguishing between human chromosomes, Nat. New Biol. 31 (1971) 232. [6] P. Balicek, J. Zizka, H. Skalska´, Variability and familial transmission of constitutive heterochromation of human chromosomes evaluated by the method of linear measurement, Hum. Genet. 42 (1978) 257 – 265. ¨ rztl. Lab. 25 [7] E. Friedrich-Scho¨ler, J. Herbich, J. Szilva´ssy, Zur zytogenetichen Vaterschaftsdiagnose, A (1979) 299 – 303. [8] G. Bujdoso´, J. Arnold, in: J. Ludwigshafen/Rh (Ed.), Genetischer Atlas der Anthropologie und Erbbiologie, GENLAB, Wu¨rzburg, Budapest, 1999, pp. 26 – 44. [9] A.J. Jeffreys, Genetic typing and confirmation of parenthood, Lancet 2 (8562) (1987 Oct. 3) 806. [10] A. La´szik, B. Brinkmann, P. So´tonyi, A. Falus, Automated fluorescent detection of a 10 loci multiplex for paternity testing, Acta Biol. Hung. 51 (1) (2000) 97 – 103.
Family relationship proven by chromosomal and DNA examinations Gyo¨rgyi Bujdoso´ a,b,*, Pe´ter So´tonyi a,b, Andras Laszik a, Max P. Baur c a
Institute of Forensic Medicine, Semmelweis University, Budapest, Hungary b Research Unit Sponsored by Hungarian Academy of Science, Hungary c Institute of Medical Biometry and Epidemiology, University of Bonn, Bonn, Germany
Abstract As of today in Hungary, the live-birth rate is continuously declining, but the number of babies born from out of marriage relationship is increasing. According to the National Statistical Office, the later number represents 28% in 1999. In the past 30 years, we have been engaged in many cases where an adult wants to be informed about his or her ancestry. Out of these plentiful caseloads, we would like to present three interesting cases. Case 1: Identification of two stepbrothers with chromosomal and DNA investigations—without examining the parents. Case 2: Proved family relationship (father – son) with chromosomal translocation studies (22/Y). Case 3: After 55 years, family members were identified (blood groups, anthropology, chromosomes, DNA) of a prisoner of war (POW). D 2003 Elsevier B.V. All rights reserved. Keywords: Hereditary; Anthropology; Chromosome; DNA; Expert
1. Material and methods In our institution, we have access to perform blood group, anthropological, chromosomal and DNA examinations. In case of anthropological examination, the persons involved will be compared using characteristic features, with ample photo documentation [1,2]. Fourteen blood and serological groups were investigated. The chromosomes were cultivated from peripheral blood after 72 h. The applied staining methods were: Q and C
* Corresponding author. Institute of Forensic Medicine, Semmelweis University, Budapest, Hungary. 0531-5131/ D 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0531-5131(03)01053-7
S20
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
banding, rarely G banding if structural chromosomal abnormalities were detected [3– 8]. The DNA studies were done according to the recommendations of International Forensic Haemogenetic Society [9,10].
2. Case report 2.1. Case 1 V.M., a 31-year-old male, who was born out of wedlock, requested an examination on his genetic relationship to his so-called stepbrother. He had never known his father and he was well in his adulthood, when he found out about M.S., who was a 28-yearold man, born from a marriage. They speculated that they may have a common father. The hypothetical father has passed away, the mother did not give her consent to the examination; therefore, we could examine only the two assumed stepbrothers. During the chromosomal studies, we highlighted the chromosome 1, 9, 16 and Y with C banding and the chromosome Y with Q banding as well. The chromosomal studies showed V.M.’s assumption to be true, which was assured by DNA examinations (Table 1). 2.2. Case 2 In this case, an older man (whose name is the same as his son’s) wants to be sure that his offspring is really his son, because 50 years ago only fingerprint examination was possible, so he was in doubt in all his adult life about being the biological father. If this is proved, he can leave him a big share in his will. The mother of this son was not available, but it was not necessary, because the chromosome 22 was translocated to the chromosome Y in both tested persons. Besides that the paternity was proved absolutely, this case represents a rarity in the scientific literature. This case will be published in the near future with more minute details (Fig. 1). Table 1 DNA analysis PCR system
M.S.
V.M.
HUMTHO1 HUMvWF D18S51 FGA D8S1179 D3S1359 D21S11 SE33
6/6 14/16 12/15 22/22 15/15 5/14 29/30 17/26.2
6/9 14/18 16/19 19/21 13/15 5/14 29/33.2 26.2/28.2
DYS 19 DYS 390 DYS 392
16 24 11
16 24 11
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
Fig. 1.
Fig. 2.
S21
S22
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
Fig. 3.
Fig. 4.
G. Bujdoso´ et al. / International Congress Series 1239 (2003) S19–S23
S23
2.3. Case 3 In this third case, the presumed family members of a lost POW were found. Anthropological, blood group, chromosomal and DNA examinations were done on the POW (A.T.) as well as on his assumed brother and sister. A.T. become a prisoner in Russia during the WW II and has spent 55 years in a camp in a lunatic asylum. The persons investigated were: Andra´s Toma, suspected half-brother and sister, Ja´nos and Anna, from paternal side. You can see on these anthropological characters: the colour of the eyes, the upper eyelid’s hottentote wrinkles, the shape of the chin and its dimple at all three persons are the same. At Andra´s and Ja´nos, the shape of the eyebrows is the same. At Andra´s and Anna, the redness of the lower lip is the same. And now you can see their father (Fig. 2). At the chromosomal investigation, the size and the intensity of fluorescence at the Ychromosome were the same at both so we learned that they got a common father. Besides that, the choromosome numbers 4, 13, 14 and 21 were the same in all three of them (Figs. 3 and 4). The parallel DNA investigations proved and supported these findings. Biostatistical results PCR systems RFLP systems Y systems Blood serology In sum
99.98% 75.64% 97.08% 59.51% 99.9999%
References [1] G. Beitzke, H. Hosemann, P. Dahr, H. Schade, Vaterschaftsgutachten fu¨r die gerinchtliche Praxis, Verlag Otto´ Schwrtz et CO, Go¨ttingen, vol. II, 1978, pp. 58 – 59. [2] M. Bujdoso´ Gy, Anthropomorph-genetika Album, Kiege´szı´te´s az egyetemi jegyzethez, Rro Renovanda Cultura Hungeriae, Budapest, 1995. [3] T. Caspersson, L. Zech, C. Johannson, Quinacrine mustardfluorescence of human chromosomes, Exp. Cell Res. 61 (1970) 474 – 475. [4] P.P. Craig-Holmes, M. Shaw, Polymorphism of human constitutive heterochromatin, Science 1974 (1971) 702 – 704. [5] A.T. Summer, H.J. Evans, R. Buckland, New technique for distinguishing between human chromosomes, Nat. New Biol. 31 (1971) 232. [6] P. Balicek, J. Zizka, H. Skalska´, Variability and familial transmission of constitutive heterochromation of human chromosomes evaluated by the method of linear measurement, Hum. Genet. 42 (1978) 257 – 265. ¨ rztl. Lab. 25 [7] E. Friedrich-Scho¨ler, J. Herbich, J. Szilva´ssy, Zur zytogenetichen Vaterschaftsdiagnose, A (1979) 299 – 303. [8] G. Bujdoso´, J. Arnold, in: J. Ludwigshafen/Rh (Ed.), Genetischer Atlas der Anthropologie und Erbbiologie, GENLAB, Wu¨rzburg, Budapest, 1999, pp. 26 – 44. [9] A.J. Jeffreys, Genetic typing and confirmation of parenthood, Lancet 2 (8562) (1987 Oct. 3) 806. [10] A. La´szik, B. Brinkmann, P. So´tonyi, A. Falus, Automated fluorescent detection of a 10 loci multiplex for paternity testing, Acta Biol. Hung. 51 (1) (2000) 97 – 103.