The next generation of DNA profiling--STR typing by multiplexed PCR--ion-pair RP LC-ESI time-of-flight MS

Pitterl,F.; Niederstätter,H.; Huber,G.; Zimmermann,B.; Oberacher,H.; Parson,W.; For the first time a multiplexed PCR approach suitable for mass spectrometric STR allele identification is presented. Thirteen forensically important STR markers (vWA, D21S11, D3S1358, D16S539, D8S1179, D7S820, D13S317, D5S818, TPOX, CSF1PO, D2S441, D10S1248, and D22S1045) and the gender typing locus amelogenin were simultaneously amplified. Ion-pair reversed-phase high-performance liquid chromatography electrospray-ionization time-of-flight mass spectrometry (ICEMS) was applied for genotyping, and allowed for highly efficient characterization of multiple PCR amplicons. [Read More]

Timing and deciphering mitochondrial DNA macro-haplogroup R0 variability in Central Europe and Middle East

Brandstätter,A.; Zimmermann,B.; Wagner,J.; Göbel,T.; Röck,A.; Salas,A.; Carracedo,A.; Parson,W.; BACKGROUND: Nearly half of the West Eurasian assemblage of human mitochondrial DNA (mtDNA) is fractioned into numerous sub-lineages of the predominant haplogroup (hg) R0. Several hypotheses have been proposed on the origin and the expansion times of some R0 sub-lineages, which were partially inconsistent with each other. Here we describe the phylogenetic structure and genetic variety of hg R0 in five European populations and one population from the Middle East. [Read More]

Unraveling the metabolic transformation of tetrazepam to diazepam with mass spectrometric methods

Schubert,B.; Pavlic,M.; Libiseller,K.; Oberacher,H.; The metabolic transformation pathways of the 1,4-benzodiazepine tetrazepam (C(16)H(17)ClN(2)O, average mass: 288.772) were studied with capillary LC-QqTOF-MS and -MS/MS by analyzing human plasma and urine samples collected from healthy volunteers. Each volunteer took 50 mg of tetrazepam, given in the form of one tablet of Myolastan (Sanofi-Synthelabo, Vienna, Austria). Accurate molecular mass measurements in full-scan mode (scan range: 50-700) were used to survey the collected samples for putative metabolic transformation products. [Read More]

Y-STR analysis on DNA mixture samples--results of a collaborative project of the ENFSI DNA Working Group

Parson,W.; Niederstätter,H.; Lindinger,A.; Gill,P.; The ENFSI (European Network of Forensic Science Institutes) DNA Working Group undertook a collaborative project on Y-STR typing of DNA mixture samples that were centrally prepared and thoroughly tested prior to the shipment. Four commercial Y-STR typing kits (Y-Filer, Applied Biosystems, Foster City, CA, USA; Argus Y Nonaplex, Biotype, Dresden, Germany; Powerplex Y, Promega, Madison, WI, USA; and DYSplex-3, SERAC, Bad Homburg, Germany) were used for the amplification of the mixture samples. [Read More]

A modular real-time PCR concept for determining the quantity and quality of human nuclear and mitochondrial DNA

Niederstätter,H.; Köchl,S.; Grubwieser,P.; Pavlic,M.; Steinlechner,M.; Parson,W.; We developed a modular real-time (rt) PCR system for absolute quantification of human nuclear (n) and mitochondrial (mt) DNA. For determination of the number of amplifiable template molecules with a minimum length required for downstream genotyping and assessment of the PCR-relevant degradation grade of the template DNA, primers yielding differently sized PCR products (nDNA: 79, 156, and 246bp; mtDNA: 102, 143, 283, and 404bp) and TaqMan hybridization probes were used for amplification and on-line product detection. [Read More]