Results from Alpha Testers

1
PowerPlex Systems for the New European Standard
Cynthia Sprecher, B.S., Bob McLaren, Ph.D., Dawn
Rabbach, A.A.S., Benjamin Krenke, M.S., Martin
Ensenberger, Ph.D., Patricia Fulmer, Ph.D., Mark
McDougall, Ph.D., Doug Storts, Ph.D.Promega
Corporation
Abstract
Robust Amplification in Presence of PCR
Inhibitors
With over 7 million profiles currently stored in
European databases, and the number expected to
increase in response to the Treaty of Prum, the
likelihood of random matches will undoubtedly
increase as well. To improve the overall power
of discrimination as well as provide
standardization across Europe, the ENFSI and
EDNAP committees have made a recommendation to
extend the current European Standard Set (ESS)
for STR systems. The PowerPlex European Systems
have been designed to comply with the new
recommendation. The kits are based on
five-color-technology and allow co-amplification
and detection of the current commonly tested
loci, plus the 5 new recommended loci. These kits
will be offered in multiple formats, including
the option to detect SE33, to accommodate various
requirements and/or preferences. The kits have
increased tolerance to common inhibitors and
increased sensitivity to obtain full profiles
from low-level DNA . Additionally, they are
robust enough to genotype degraded DNA samples
through the use of mini STR loci. Product launch
is expected in Fall 2009.
PowerPlex ESX 170.5ng DNA plus200mM Hematin
Introduction
The 5 new STR loci recommended by ENFSI (D2S441,
D10S1248, D22S1045, D1S1656, and D12S391) were
chosen primarily due to the small sizes of their
amplicons, which are considered more useful when
working with degraded DNA samples. ENFSI
requested that vendors make available prototype
STR multiplexes containing the 11 SGM Plus loci
and the 5 ENFSI loci by January 2009 for
testing. Promega has developed a new 16 locus
multiplex to meet this goal (PowerPlex ESX 16).
In response to feedback from key European labs we
are also developing a slightly different 16 locus
multiplex where the SGM Plus loci have been made
smaller and the new ENFSI loci are larger
(PowerPlex ESI 16). Using both kit
configurations, one may amplify up to 15 loci
with amplicons lt250 bases (including 10 of the
11 SGM Plus loci). This will enable recovery of
more information from degraded DNA samples. A
second 17-plex version of each of these
multiplexes will be made to include the SE33
locus for those who choose to test for this
locus.
D2S441
D12S391
D19S433
SE33
PowerPlex ESI 17 0.5ng DNA plus60ng/uL Humic
Acid
PowerPlex ESX and PowerPlex ESI Loci
Arrangements
Results from Alpha Testers
PowerPlex ESX 17
New ENFSI loci present as miniSTR amplicons
(circled). Profile is from 0.5ng human gDNA
amplified for 30 cycles
Degraded DNA
PowerPlex ESX 17 Amp of 1.5ng DNA from 70 Year
OldBone Sample
Data courtesy ofP. Martin, C. Fernandez, and A.
Alonso, Facultatives Biology Service-
INTCF,Instituto Nacional de Toxicologia y
Ciencias Forenses, Spain
PowerPlex ESI 17
D1S1656
D2S441
New ENFSI loci present as larger STR amplicons
(circled) Profile is from 0.5ng human gDNA
amplified for 30 cycles
Sensitivity
D12S391
PowerPlex ESI 17 Amp of 62.5pg DNA
Data courtesy ofA. Berti, Carabinieri Scientific
Dept of Rome,Italy
D8S1179
FGA
SE33
Concordance Study
Acknowledgements
We would like to extend a special thank you to
John Butler, Becky Hill, Margaret Kline, and
David L. Duewer from NIST for their time and
effort in performing the concordance study.
After addition of add-in primer to account for a
G to T SNP in primer binding site at D22S1045
locus in PowerPlex ESX configuration. This SNP
was seen in 4 samples.
SGM Plus is a registered trademark of Life
Technologies Corporation or its affiliates in the
US and/or certain other countries