Speziesidentifizierung mittels vergleichender Sequenzanalyse des mitochondrialen 12S-rRNA-Gens

RNA (rRNA), die eine wichtige, primär funktionale Rolle in der Zellphysiologie einnimmt. Die Gene dieser rRNA lassen sich in hochkonservierte und hochpolymorphe Bereiche unterscheiden. Während in den konservierten Regionen kaum Mutationen zu finden sind, werden in den polymorphen Abschnitten je nach Grad der Verwandtschaft große Sequenzunterschiede zwischen den verschiedenen Tierarten beobachtet. Individuen einer Art zeigen diese Unterschiede nicht. Somit können diese artspezifischen Abweichungen dazu genutzt werden, biologische Materialien unbekannter Herkunft einer bestimmten Tierart zuzuordnen. Mit der Amplifikation und anschließenden Sequenzierung eines Bereiches innerhalb des mitochondrialen 12S-Gens sowie der Auswertung bereits publizierter Sequenzen gelingt die Identifizierung einer Spezies anhand von 20 – 25 Basen. Dazu kann neben der etablierten Methode der Sequenzierung nach Sanger auch die Technik der Pyrosequenzierung genutzt werden. Die Ergebnisse zeigen die Möglichkeit der Identifizierung verschiedener Arten durch die Analyse kurzer Fragmente ihrer 12S-Gen-Sequenz. Dazu reicht die Amplifikation des gewünschten Fragmentes mit Hilfe eines Primerpaares. Für degradierte DNA wurde ein alternativer Rückprimer getestet, der die Amplifikation eines kürzeren Sequenzabschnittes ermöglicht. Es wurden insgesamt 91 Proben verschiedener Tiere analysiert, die sich aus 8 Säugetier-Arten, 2 Fisch-Arten und 3 Vogel-Arten zusammen setzten. Zudem kann und wird die Methode bereits bei Fragestellungen in der forensischen Routine angewendet

Allelic proportions of 16 STR loci—including the new European Standard Set (ESS) loci—in a Swiss population sample

Allele frequencies and forensically relevant population statistics of 16 STR loci, including the new European Standard Set (ESS) loci, were estimated from 668 unrelated individuals of Caucasian appearance living in different parts of Switzerland. The samples were amplified with a combination of the following three kits: AmpFlSTR® NGM SElect™, PowerPlex® ESI17 and PowerPlex® ESX 17. All loci were highly polymorphic and no significant departure from Hardy-Weinberg equilibrium and linkage equilibrium was detected after correction for sampling

A global analysis of Y-chromosomal haplotype diversity for 23 STR loci

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent.Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marino, Miguel Eduardo. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Analisis de ADN; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Purps, Josephine. Charité-Universitätsmedizin; AlemaniaFil: Siegert, Sabine. University of Cologne; AlemaniaFil: Willuweit, Sascha. Charité-Universitätsmedizin; AlemaniaFil: Nagy, Marion. Charité-Universitätsmedizin; AlemaniaFil: Alves, Cíntia. Universidad de Porto; PortugalFil: Salazar, Renato. Universidad de Porto; PortugalFil: Angustia, Sheila M. T.. Philippine National Police Crime Laboratory; FilipinasFil: Santos, Lorna H.. Philippine National Police Crime Laboratory; FilipinasFil: Anslinger, Katja. Universitat Genzentrum Der Ludwing-maximilians; AlemaniaFil: Bayer, Birgit. Universitat Genzentrum Der Ludwing-maximilians; AlemaniaFil: Ayub, Qasim. The Wellcome Trust Sanger Institute; Reino UnidoFil: Wei, Wei. The Wellcome Trust Sanger Institute; Reino UnidoFil: Xue, Yali. The Wellcome Trust Sanger Institute; Reino UnidoFil: Tyler Smith, Chris. The Wellcome Trust Sanger Institute; Reino UnidoFil: Baeta Bafalluy, Miriam. Universidad de Zaragoza; EspañaFil: Martínez Jarreta, Begoña. Universidad de Zaragoza; EspañaFil: Egyed, Balazs. Eotvos University, Budapest; ArgentinaFil: Balitzki, Beate. Universidad de Basilea; SuizaFil: Tschumi, Sibylle. Universidad de Basilea; SuizaFil: Ballard, David. King; Reino UnidoFil: Syndercombe Court, Denise. King; Reino UnidoFil: Barrantes, Xinia. Poder Judicial, Forensic Sciences Department; Costa RicaFil: Bäßler, Gerhard. Landeskriminalamt Baden-Württemberg; AlemaniaFil: Berger, Burkhard. Universidad de Innsbruck; AustriaFil: Niederstätter, Haral. Universidad de Innsbruck; AustriaFil: Parson, Walther. Universidad de Innsbruck; Austria. University Park; Estados UnidosFil: Davis, Carey. Department of Molecular and Medical Genetics; Estados Unidos. Institute of Applied Genetics; Estados UnidosFil: Furfuro, Sandra. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; ArgentinaFil: Locarno, Laura. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentin

A global analysis of Y-chromosomal haplotype diversity for 23 STR loci

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent.Peer reviewe

[Reconstruction of sexual offences--forensic aspects of sperm traces]

The investigation of sexual offences is a real challenge, as the injuries are often unspecific or faint and may sometimes be missing completely. Evidence recovery and analysis as well as the statements of the victims and suspects are therefore of vital importance. In both presented cases, the results of trace evidence analysis were basically consistent with a sexual assault, but the victims' statements regarding the course of events and the pattern of traces showed severe discrepancies

Allelic proportions of 16 STR loci—including the new European Standard Set (ESS) loci—in a Swiss population sample

Allele frequencies and forensically relevant population statistics of 16 STR loci, including the new European Standard Set (ESS) loci, were estimated from 668 unrelated individuals of Caucasian appearance living in different parts of Switzerland. The samples were amplified with a combination of the following three kits: AmpFlSTR® NGM SElect?, PowerPlex® ESI17 and PowerPlex® ESX 17. All loci were highly polymorphic and no significant departure from Hardy-Weinberg equilibrium and linkage equilibrium was detected after correction for sampling