Abstract
A multiplex assay with 49 autosomal single nucleotide polymorphisms (SNPs) developed
for human identification was validated for forensic genetic casework and accredited
according to the ISO 17025 standard. The multiplex assay was based on the SNPforID 52plex SNP assay [J.J. Sanchez, C. Phillips, C. Børsting, K. Balogh, M. Bogus,
M. Fondevila, C.D. Harrison, E. Musgrave-Brown, A. Salas, D. Syndercombe-Court, P.M.
Schneider, A. Carracedo, N. Morling, A multiplex assay with 52 single nucleotide polymorphisms
for human identification, Electrophoresis 27 (2006) 1713–1724], where 52 fragments
were amplified in one PCR reaction. The SNPs were analysed by single base extension
(SBE) and capillary electrophoresis. Twenty-three of the original SBE primers were
altered to improve the overall robustness of the assay and to simplify the analysis
of the SBE results. A total of 216 samples from 50 paternity cases and 33 twin cases
were typed at least twice for the 49 SNPs. All electropherograms were analysed independently
by two expert analysts prior to approval. Based on these results, detailed guidelines
for analysis of the SBE products were developed. With these guidelines, the peak height
ratio of a heterozygous allele call or the signal to noise ratio of a homozygous allele
call is compared with previously obtained ratios. A laboratory protocol for analysis
of SBE products was developed where allele calls with unusual ratios were highlighted
to facilitate the analysis of difficult allele calls. The guidelines for allele calling
proved to be highly efficient for the detection of DNA mixtures and contaminated DNA
preparations. DNA from two individuals was mixed in seven different ratios ranging
from 1:1 to 1:10; all mixtures were easily identified as mixtures.
Keywords
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Article info
Publication history
Published online: May 08, 2009
Accepted:
April 9,
2009
Received in revised form:
April 3,
2009
Received:
February 23,
2009
Identification
Copyright
© 2009 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
