Summary
Paternity testing using DNA polymorphism of variable numbers of tandem repeat (VNTR) regions with restriction fragment length polymorphism (RFLP) was implemented. HinfI-digested DNA was separated by electrophoresis in agarose gels and hybridized with radiolabelled probes detecting the VNTR-systems D2S44 (YNH24), D5S43 (MS8), D7S21 (MS31), D7S22 (g3), and D12S11 (MS43a). The intra gel variability of 970 duplicate investigations on the same gel of DNA from 122 individuals showed no differences exceeding 1.25 mm between the positions of the corresponding DNA fragments. The comparison of 1,624 DNA fragments from 342 mother/child pairs showed only one difference above 1.25 mm which was interpreted as a mutation. Based on these observations, we decided to consider an intra gel difference above 1.25 mm between the non-maternal DNA fragment of the child and the nearest DNA fragment of the putative father as an exclusion in paternity testing. This matching criterion was used for the comparisons of 1,197 DNA fragment differences in 247 pairs of children and putative fathers who had not been excluded by conventional marker systems. In all of these cases, the migration differences between the DNA fragments of non-excluded men and the DNA fragments of the children were less than 1.25 mm except in 6 cases (0.5%). The man/child differences in all of 227 false trios exceeded 1.25 mm in 2 or more of the 5 VNTR systems investigated. Matching criteria for inter gel comparisons in paternity testing were established. The frequency distribution of Hinfl digested DNA fragments of the 5 VNTR systems in 650 unrelated Danes is presented and the raw data is available.
Zusammenfassung
DNA-Polymorphismen mit einer variablen Anzahl von tandemähnlichen Wiederholungseinheiten (VNTR's), speziell der Typus der Restriktionsfragmentlängenpolymorphismen, wurden in die Vaterschaftsanalyse eingeführt. Hinfl-verdaute DNA wurde elektrophoretisch in Agarose-Gelen aufgetrennt und mit radioaktiv markierten Sonden hybridisiert, welche die VNTR-Systeme D2S44 (YNH24), D5S43 (MS8), D7S21 (MS31), D7S22 (G3) und D12S11 (MS43a) detektieren. Die sog. Intra-Gelvariation von 970 Doppeluntersuchungen auf demselben Gel von DNA von 122 Personen zeigte keine Unterschiede, welche größer waren als 1,25 mm — bezogen auf die Positionen der korrespondierenden DNA-Fragmente. Der Vergleich von 1.624 DNA-Fragmenten von 342 Mutter—Kind—Paaren zeigte lediglich einen Unterschied, welcher größer als 1,25 mm war und daher als eine Mutation interpretiert wurde. Hierauf basierend entschlossen wir uns, eine Intra-Geldifferenz größer als 1,25 mm zwischen dem nicht-mütterlichen DNA-Fragment des Kindes und dem nächsten DNA-Fragment des Putativ-Vaters als einen Ausschluß in der Vaterschaftsanalyse zu bewerten. Dieses Match-Kriterium wurde benutzt für die Vergleiche von 1.197 DNA-Fragmentdifferenzen bei 247 Paaren von Kindern und Putativ-Vätern, bei welchen in konventionellen Systemen kein Ausschluß zu beobachten war. In all diesen Fällen waren die Wanderungsunterschiede zwischen DNA-Fragmenten der nicht-ausgeschlossenen Männner und den Fragmenten der Kinder geringer als 1,25 mm mit der Ausnahmen von 6 Fällen (0,5%). Die Mann—Kind—Differenzen in allen der 227 falschen Terzette überschritten 1,25 mm in zwei oder mehr der 5 VNTR-Systeme, welche untersucht wurden. Match-Kriterien für die Inter-Gel-Vergleiche bei Vaterschaftsuntersuchungen wurden etabliert. Die Frequenz-Verteilung von HinfIverdauten DNA-Fragmenten der 5 VNTR-Systeme bei 650 unverwandten Dänen wird gezeigt und die Rohdaten sind verfügbar.
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Morling, N., Hansen, H.E. Paternity testing with VNTR DNA systems. Int J Leg Med 105, 189–196 (1993). https://doi.org/10.1007/BF01642792
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DOI: https://doi.org/10.1007/BF01642792