Abstract
The AmpFℓSTR SEfiler kit co-amplifies 11 short tandem repeat loci including SE33 in a single multiplex. After establishing the optimum in primer titration studies, the primer concentrations of all loci in the multiplex were chosen such that the heterozygote peak height ratios of each of the loci were balanced. The combined primer set was then tested to determine the robustness of the multiplex under various conditions. Different MgCl2 concentrations were evaluated to establish the optimum concentration for the multiplex. The amplification of the various loci in the multiplex was tested at several annealing temperatures (55–63°C). Additionally, DNA from primates, non-primates and microorganisms were amplified to investigate the specificity of the kit. The stability of the AmpFℓSTR SEfiler kit was determined by addition of hematin, to simulate inhibition, and the use of degraded DNA. Population studies revealed a probability of identity of 6.47×10−15 for African Americans and 7.46×10−14 for US Caucasians. To assess the ability of the multiplex to analyze forensic samples, testing on blood, oral swabs and mixtures was performed. Based on the various studies, it was determined that the AmpFℓSTR SEfiler PCR amplification kit can be used to successfully analyze a variety of forensic, databasing and paternity samples.
Similar content being viewed by others
References
Bäßler G, Förster R, Eberspächer BC, Karl M, Kugler M, Pflug W (1999) Frequency data for the STR loci HumFibra (FGA) and HumACTBP2 (SE33) in a population of Germans and Turks from South-West Germany. Int J Legal Med 112:136–138
Brinkmann B, Sajantila A, Goedde HW, Nishi K, Wiegand P (1996) Population genetic comparisons among eight populations using allele frequency and sequence data from three microsatellite loci. Eur J Hum Genet 4:175–182
Budowle B, Collins PJ, Dimsoski P et al. (2001) Population data on the STR loci D2S1338 and D19S433. Forensic Science Comm 3:1–4
Corte-Real F, Andrade L, Vieira DN, Carracedo A, Vide MC (1999) Allele distribution of three STR’s in a population from Mozambique. Int J Legal Med 112:345–347
Dimo-Simonin N, Grange F, Kratzer A, Brandt-Casadevall C, Mangin P (1998) Forensic validation of the short tandem repeat HUMACTBP2 using capillary electrophoresis. Electrophoresis 19:256–261.
DNA Advisory Board (2000) Quality assurance standards for forensic DNA testing laboratories. Forensic Sci Comm2
Gill P, Brinkmann B, d’Aloja E et al. (1997) Considerations from the European DNA profiling group (EDNAP) concerning STR nomenclature. Forensic Sci Int 87:185–192
Gill P, Kimpton C, D’Aloja E et al. (1994) Report of the European DNA profiling group (EDNAP)—towards standardization of short tandem repeat (STR) loci. Forensic Sci Int 65:51–59
Hering S, Edelmann J, Dressler J (2002) Sequence variations in the primer binding regions of the highly polymorphic STR system SE33. Int J Legal Med 116:365–367
Hochmeister MN, Rudin O, Ambach E (1998) PCR analysis from cigarette butts, postage stamps, envelope sealing flaps and other saliva-stained material. Methods Mol Biol 98:27–32
Holt CL, Buoncristiani M, Wallin JM, Nguyen T, Lazaruk KD, Walsh PS (2002) TWGDAM validation of AmpFℓSTR® PCR amplification for forensic DNA casework. J Forensic Sci 47:15–45
Junge A, Lederer T, Braunschweiger G, Madea B (2003) Validation of the multiplex kit genRESMPX-2 for forensic casework analysis. Int J Legal Med 117:317–325
Klintschar M, Neuhuber F (1998) A study of the short tandem repeat system ACTBP2 (SE33) in an Austrian population sample. Intl J Legal Med 111:46–48
Lászik A, Sótonyi P, Rand S, Hohoff C (2001) Frequency data for the STR locus ACTBP2 (SE33) in eight populations. Int J Legal Med 115:94–96
Lazaruk K, Wallin J, Holt C, Nguyen T, Walsh PS (2001) Sequence variation in human and other primates at six short tandem repeat loci used in forensic identity testing. Forensic Sci Int 119:1–10
Lessig R, Edelmann J, Akkad Y (2000) Population genetics of ACTBP2 (SE-33) in Western Saxony (Germany). Forensic Sci Int 113:39–41
Liu C, Harashima N, Katsuyama Y, Ota M, Arakura A, Fukushima H (1997) ACTBP2 gene frequency distribution and sequencing of the allelic ladder and variants in the Japanese and Chinese populations. Int J Legal Med 110:208–212
Möller A, Schürenkamp M, Brinkmann B (1995) Evaluation of an ACTBP2 ladder composed of 26 sequenced alleles. Int J Legal Med 108:75–78
Pestoni C, Lareu MV, López-Gómez J, Carracedo A (1999) Genetic data on three complex STRs (ACTBP2, D21S11, and HUMFIBRA/FGA) in the Galician population (NW Spain). Int J Legal Med 112:337–339
Rand S, Schürenkamp M, Brinkamnn B (2002) The GEDNAP (German DNA profiling group) blind trial concept. Int J Legal Med 116:199–206
Rolf B, Schürenkamp M, Junge A, Brinkmann B (1997) Sequence polymorphism at the tetranucleotide repeat of the human beta-actin related pseudogene H-beta-Ac-psi-2 (ACTBP2) locus. Int J Legal Med 110:69–72
Rothämel T, Kleemann WJ, Tröger HD (2000) Reliability of SE33 typing by capillary electrophoresis. Forensic Sci Int 113:33–38
Schmitter H, Sonntag M-L (1995) STR-analysis on cigarette-butts: experiences with casework material. Klin Lab 41:177–180
Schneider HR, Rand S, Schmitter H, Weichhold G (1998) ACTBP2-nomenclature recommendations of GEDNAP. Int J Legal Med 111:97–100
Shimada I, Rand S, Brinkmann B, Hohoff C (2002) Kurdish population data from 11 STR loci (ACTBP2, CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D13S317 and D21S11) Int J Legal Med 116:301–303
Sparkes R, Kimpton C, Gilbard S, Carne P, Anderson J, Oldroyd N, Urquhart A, Gill P (1996) The validation of a 7-locus multiplex STR test for use in forensic casework. (II) Artefacts, casework studies and success rates. Int J Legal Med 109:195–204
Van Hoofstat D, Deforce D, Millecamps R et al. (1998) Population genetic study of four short tandem repeat loci in the Belgian population, using capillary electrophoresis. Electrophoresis 19:719–722
Wallin J, Holt CL, Lazaruk KD, Nguyen T, Walsh PS (2002) Constructing universal multiplex PCR systems for comparative genotyping. J Forensic Sci 47:1–14
Wallin JM, Buoncristiani MR, Lazaruk KD, Fildes N, Holt CL, Walsh S (1998) TWGDAM validation of the AmpFℓSTR® Blue PCR amplification kit for forensic casework analysis. J Forensic Sci 43:854–870
Wiegand P, Budowle B, Rand S, Brinkman B (1993) Forensic validation of the STR systems SE33 and TC11. Int J Legal Med 105:315–320
Acknowledgments
The authors would like to thank the members of the Human Identification group at Applied Biosystems for their comments. The authors would specifically like to thank Dr. Dennis Reeder for his useful suggestions.
Author information
Authors and Affiliations
Corresponding author
Additional information
For research, forensic or paternity use only. Not for use in diagnostic procedures.
The PCR process is covered by patents owned by Roche Molecular Systems, Inc. and F. Hoffmann-La Roche Ltd.
Applied Biosystems, ABI PRISM, AmpFℓSTR, GeneScan, Genotyper, LIZ, MicroAmp, SGM Plus, and VIC are registered trademarks and FAM, Hi-Di, Identifiler, NED, PET, POP, POP-4, and ROX are trademarks of Applera Corporation or its subsidiaries in the US and certain other countries.
AmpliTaq Gold, GeneAmp, and QuantiBlot are registered trademarks of Roche Molecular Systems, Inc.
Rights and permissions
About this article
Cite this article
Coticone, S.R., Oldroyd, N., Philips, H. et al. Development of the AmpFℓSTR SEfiler PCR amplification kit: a new multiplex containing the highly discriminating ACTBP2 (SE33) locus. Int J Legal Med 118, 224–234 (2004). https://doi.org/10.1007/s00414-004-0459-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00414-004-0459-y