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Inter-laboratory study on standardized MPS libraries: evaluation of performance, concordance, and sensitivity using mixtures and degraded DNA
Journal article   Open access   Peer reviewed

Inter-laboratory study on standardized MPS libraries: evaluation of performance, concordance, and sensitivity using mixtures and degraded DNA

Petra Müller, Christian Sell, Thorsten Hadrys, Johannes Hedman, Steffi Bredemeyer, Francois-Xavier Laurent, Lutz Roewer, Sabrina Achtruth, Maja Sidstedt, Titia Sijen, …
International journal of legal medicine, v 134(1), pp 185-198
Jan 2020
DOI: https://doi.org/10.1007/s00414-019-02201-2
PMID: 31745634
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://link.springer.com/content/pdf/10.1007/s00414-019-02201-2.pdfView
Published, Version of Record (VoR) Open
url
https://doi.org/10.1007/s00414-019-02201-2View
Published, Version of Record (VoR) Open

Abstract

Alleles Austria DNA Fingerprinting - methods Electrophoresis, Capillary Female France Gene Library Germany High-Throughput Nucleotide Sequencing Humans Laboratories Male Microsatellite Repeats Netherlands Polymerase Chain Reaction Polymorphism, Single Nucleotide Sensitivity and Specificity Sequence Analysis, DNA Sweden
We present results from an inter-laboratory massively parallel sequencing (MPS) study in the framework of the SeqForSTRs project to evaluate forensically relevant parameters, such as performance, concordance, and sensitivity, using a standardized sequencing library including reference material, mixtures, and ancient DNA samples. The standardized library was prepared using the ForenSeq DNA Signature Prep Kit (primer mix A). The library was shared between eight European laboratories located in Austria, France, Germany, The Netherlands, and Sweden to perform MPS on their particular MiSeq FGx sequencers. Despite variation in performance between sequencing runs, all laboratories obtained quality metrics that fell within the manufacturer's recommended ranges. Furthermore, differences in locus coverage did not inevitably adversely affect heterozygous balance. Inter-laboratory concordance showed 100% concordant genotypes for the included autosomal and Y-STRs, and still, X-STR concordance exceeded 83%. The exclusive reasons for X-STR discordances were drop-outs at DXS10103. Sensitivity experiments demonstrated that correct allele calling varied between sequencing instruments in particular for lower DNA amounts (≤ 125 pg). The analysis of compromised DNA samples showed the drop-out of one sample (FA10013B01A) while for the remaining three degraded DNA samples MPS was able to successfully type ≥ 87% of all aSTRs, ≥ 78% of all Y-STRs, ≥ 68% of all X-STRs, and ≥ 92% of all iSNPs demonstrating that MPS is a promising tool for human identity testing, which in return, has to undergo rigorous in-house validation before it can be implemented into forensic routine casework.

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19 citations in Web of Science
21 citations in Scopus

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Collaboration types
Domestic collaboration
International collaboration
Web of Science research areas
Medicine, Legal
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