As solid as a rock—comparison of CE- and MPS-based analyses of the petrosal bone as a source of DNA for forensic identification of challenging cranial bones

Kulstein, Galina; Hadrys, Thorsten; Wiegand, Peter

doi:10.1007/s00414-017-1653-z

Cited by 41 publications

(26 citation statements)

References 31 publications

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“…In general, our results support the findings of earlier publications [74][75][76] that the application of MPS for STR analysis of degraded and challenging DNA samples can be beneficial. As expected and due to the nature of SNP-containing amplicons [25,26], the ancient DNA data showed that, in contrast to STRs, the majority of sequencing runs obtained constantly high SNP recovery rates, which was in line with previous reports [8,75] ( Table 3).…”

Section: Ancient Dna Samplessupporting

confidence: 91%

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

et al. 2019

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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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Section: Ancient Dna Samplessupporting

confidence: 91%

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

et al. 2019

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“…Recommendations have recently been made for the anthropologist to remove the otic capsule during initial examination and submitting the powder to the DNA laboratory . The otic capsule is ossified during gestation and is considered the densest part of the petrous .…”

Section: Discussionmentioning

confidence: 99%

“…In order to remove the otic capsule without removing the temporal, the base of the skull just over the otic capsule may be shaved or ground away until the capsule can be seen. The capsule may be sanded with a rasp and the powder collected through the foramen magnum or ground out with a drill bit .…”

Section: Discussionmentioning

confidence: 99%

Getting Ahead: Extraction of DNA from Skeletonized Cranial Material and Teeth

Edson

2019

Journal of Forensic Sciences

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Between 1990 and 2018, the Defense POW/MIA Accounting Agency submitted 2177 cranial elements and 1565 teeth to the Armed Forces Medical Examiner System—Armed Forces DNA Identification Laboratory for DNA testing. In an effort to identify missing United States service members, materials were recovered from wartime losses inclusive of World War II, the Korean War, and Southeast Asia. Using four different DNA extraction protocols, DNA testing was performed using mitochondrial DNA Sanger sequencing, modified AmpFlSTR® Yfiler™, AmpFlSTR® MiniFiler™, PowerPlex® Fusion, or Next Generation Sequencing. This paper aims to provide optimal strategies for the DNA testing of skeletonized cranial materials. Cranial elements produced the most consistent results in Sanger sequencing using an organic purification; however, teeth were most successful for the same platform with an inorganic purification. The inverse is true for STR testing of cranial bones. Of the cranial elements, the temporal provided the most consistent results.

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“…The International Committee for Missing Persons (ICMP) recommends an ordered sampling strategy to begin with the femur and proceeding through the long bones as has the Armed Forces Medical Examiner System–Armed Forces DNA Identification Laboratory (AFMES‐AFDIL) . More recent studies recommend the removal of the petrous portion of the cranium , believed to be the densest bone in the body.…”

mentioning

confidence: 99%

Extraction of DNA from Skeletonized Postcranial Remains: A Discussion of Protocols and Testing Modalities

Edson

2019

Journal of Forensic Sciences

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This paper provides a retrospective of the DNA analysis performed by the Armed Forces Medical Examiner–Armed Forces DNA Identification Laboratory between 1990 and 2018. Over 13,000 postcranial osseous materials, comprised of wartime losses from World War II, the Korean War, and South‐East Asia, were examined by the following: mitochondrial DNA sequencing, a modified AmpFlSTR® Yfiler™, AmpFlSTR® MiniFiler™, PowerPlex® Fusion, or NGS. Four different DNA extraction protocols were used: incomplete demineralization coupled with an organic purification; complete demineralization with an organic purification; complete demineralization with an inorganic purification using QIAquick PCR Purification Kit; and a protocol designed specifically for use with next‐generation sequencing. In general, complete demineralization coupled with an organic purification was the optimal extraction protocol for sequencing of mitochondrial DNA, regardless of the osseous element tested. For STR testing, demineralization paired with an inorganic purification provided optimum results, regardless of kit used or osseous element tested.

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As solid as a rock—comparison of CE- and MPS-based analyses of the petrosal bone as a source of DNA for forensic identification of challenging cranial bones

Cited by 41 publications

References 31 publications

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

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

Getting Ahead: Extraction of DNA from Skeletonized Cranial Material and Teeth

Extraction of DNA from Skeletonized Postcranial Remains: A Discussion of Protocols and Testing Modalities

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