Casework testing of the multiplex kits AmpFℓSTR® SEfiler Plus™ PCR amplification kit (AB), PowerPlex® S5 System (Promega) and AmpFℓSTR® MiniFiler™ PCR

Abstract

The STR kits SEfiler Plus™ (D3S1358, FGA, D8S1179, D18S51, D21S11, TH01, VWA, SE33, D2S1338, D16S539, D19S433, and Amelogenin), PowerPlex® S5 System (D18S51, D8S1179, TH01, FGA, Amelogenin) and MiniFiler™ (D13S317, D7S820, Amelogenin, D2S1338, D21S11, D16S539, D18S51, CSF1PO, and FGA) were comparatively tested for their robustness and sensitivity. About fifty stains with highly degraded DNA and little DNA quantity served as examination material (e.g. hair with a telogen root, bones, degraded saliva stains on drinking-vessels and skin cell mixtures).

The PowerPlex® S5 with five German-DNA-database (DAD)-systems and the MiniFiler kit with four topical DAD-systems and further STR markers show reduced amplicon lengths. The SEfiler Plus™ kit represents no MiniSTR multiplex, but contains the nine current DAD-systems and further three systems D2S1338, D16S539, D19S433, which are the potential expansion markers for the German-DNA-database.

We have detected on the basis of our comparative stain investigations, that the SEfiler Plus kit was less sensitive than the PowerPlex® S5 and the MiniFiler kit. The MiniFiler™ and the PowerPlex® S5 kit showed a comparative high sensitivity.

Especially in analysing skin cell mixtures, the MiniFiler kit contained larger differences with regard to the performance of the fluorescent dyes/primer concentration co-ordination than the PowerPlex® S5. The SEfiler Plus kit generated – just as both MiniSTR kits – relative robust typing results, but there appeared an increased sensitivity for “allelic drop-outs” and “imbalances”. Since the SEfiler Plus kit was not planned as MiniSTR concept, “allelic drop-outs” were observed, as expected, more frequent in typing stains with degraded DNA and little DNA quantity, especially in the long PCR products (e.g. D18S51).

Origine: fsigenetics.com