Inter-laboratory study on standardized MPS libraries: evaluation of performance, concordance, and sensitivity using mixtures and degraded DNA
| Autor: | Sascha Willuweit, Thorsten Hadrys, Lutz Roewer, Steffi Bredemeyer, Titia Sijen, Natalie E.C. Weiler, Sabrina Achtruth, Marc Trimborn, Ingo Bastisch, Christian Sell, F.-X. Laurent, Petra Müller, Walther Parson, Johannes Hedman, Maja Sidstedt |
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| Rok vydání: | 2019 |
| Předmět: |
Male
Massively parallel sequencing Concordance Method Paper Locus (genetics) Computational biology Biology Polymerase Chain Reaction Polymorphism Single Nucleotide Sensitivity and Specificity Pathology and Forensic Medicine 03 medical and health sciences 0302 clinical medicine Germany Genotype Humans 030216 legal & forensic medicine Inter-laboratory Collaborative study Alleles 030304 developmental biology Gene Library Netherlands Sweden 0303 health sciences Massive parallel sequencing Electrophoresis Capillary High-Throughput Nucleotide Sequencing Sequence Analysis DNA DNA Fingerprinting Ancient DNA Short tandem repeats Austria Microsatellite Female Degraded dna France Laboratories Inter-laboratory study ForenSeq DNA Signature Prep Kit Microsatellite Repeats |
| Zdroj: | International Journal of Legal Medicine |
| ISSN: | 1437-1596 |
| Popis: | 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. Electronic supplementary material The online version of this article (10.1007/s00414-019-02201-2) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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