DNA typing from skeletal remains:: Evaluation of multiplex and megaplex STR systems on DNA isolated from bone and teeth samples

Aim. To evaluate the performance of three multiplex short tandem repeat (STR) systems (Ampf/STR Profiler (TM), Ampf/STR Profiler PIus (TM), and Ampf/STR COfiler (TM)), and a megaplex STR system (PowerPlex (TM) 1 6) on DNA extracted from the skeletal remains. By performing a microbial DNA challenge study, we also evaluated the influence of microbial DNA on human DNA typing. Methods. A subset of 86 DNA extracts isolated from 8-50 years old bone and teeth samples, corresponding to 20 identification cases from mass graves in CI oatia and Bosnia and Herzegovina, and to 4 paternity cases involving deceased parents in Spain, were analyzed by the above systen7s. Results. Bone samples with no detectable human DNA (tested with Quantiblot), as well as teeth samples with deteetable human DNA, were successfully amplified. Surprisingly, even in highly degraded samples, PowerPlex (TM) 16 offered very robust amplification for the both Penta E and Penta D markers. We observed a few non-specific extra peaks of 202 and 308 base pairs, which appeared to match 16S rRNA of the Pseudomonas halodenitrificans. Conclusion. Ampf/STR Profiler (TM) Kit, Ampf/STR Profiler plus (TM) Kit, the Ampf/STR COfiler (TM) Kit, and the PowerPlex (TM) 16 system are very sensitive multiplex STR amplification systems, which can be successfully used to obtain a multilocus STR profile from old teeth and bone samples with minimal amounts (pg) of human DNA or even with no detectable human DNA.