Which tissue to take? A retrospective study of the identification success of altered human remains

Senst, Alina; Scheurer, Eva; Gerlach, Kathrin; Schulz, Iris

doi:10.1016/j.jflm.2021.102271

Cited by 7 publications

(10 citation statements)

References 22 publications

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“…In particular, buccal swabs extracted with the SwabSolution™ Kit and amplified with DPMB showed the most unpredictable number of reads and the lowest genotyping success. Such a deviation was also seen in a previous work [28], but the underlying samples were from altered and degraded human materials, for which divergent results are expected. Potentially different sequencing efficiencies could cause a general poor agreement in read intensities despite equal DNA input from fresh samples, which were found in the repetition and reproduction studies.…”

Section: Discussionsupporting

confidence: 70%

“…previous work [28], but the underlying samples were from altered and degraded human materials, for which divergent results are expected. Potentially different sequencing efficiencies could cause a general poor agreement in read intensities despite equal DNA input from fresh samples, which were found in the repetition and reproduction studies.…”

Section: Variancesmentioning

confidence: 89%

“…An insufficient amount of DNA provokes an increase in the formation of adapter dimers, which can negatively impact the sequencing quality [26,27]. Especially DNA from postmortem tissue samples of altered corpses can be highly degraded and inhibited due to the decomposition process [18,28]. Identifying human remains with high postmortem intervals is a common task in forensic medicine, yet NGS technologies for STR and SNP analysis of altered tissue samples has not been widely evaluated [4].…”

Section: Introductionmentioning

confidence: 99%

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Validation and beyond: Next generation sequencing of forensic casework samples including challenging tissue samples from altered human corpses using the MiSeq FGx system

Senst¹,

Caliebe²,

Scheurer³

et al. 2022

Journal of Forensic Sciences

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The proceeding developments in next generation sequencing (NGS) technologies enable increasing discrimination power for short tandem repeat (STR) analyses and provide new possibilities for human identification. Therefore, the growing relevance and demand in forensic casework display the need for reliable validation studies and experiences with challenging DNA samples. The presented validation of the MiSeq FGx system and the ForenSeq™ DNA Signature Prep Kit (1) investigated sensitivity, repeatability, reproducibility, concordance, pooling variations, DNA extraction method variances, DNA mixtures, degraded, and casework samples and (2) optimized the sequencing workflow for challenging samples from human corpses by testing additional PCR purification, pooling adjustments, and adapter volume reductions. Overall results indicate the system's reliability in concordance to traditional capillary electrophoresis (CE)‐based genotyping and reproducibility of sequencing data. Genotyping success rates of 100% were obtained down to 62.5 pg DNA input concentrations. Autosomal STR (aSTR) profiles of artificially degraded samples revealed significantly lower numbers of locus and allelic dropouts than CE. However, it was observed that the system still exposed drawbacks when sequencing highly degraded and inhibited samples from human remains. Due to the lack of studies evaluating the sequencing success of samples from decomposed or skeletonised corpses, the presented optimisation studies provide valuable recommendations such as an additional PCR purification, an increase in library pooling volumes, and a reduction of adapter volumes for samples with concentrations ≥31.2 pg. Thus, this research highlights the importance of all‐encompassing validation studies for implementing novel technologies in forensic casework and presents recommendations for challenging samples.

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Section: Discussionsupporting

confidence: 70%

Section: Variancesmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

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Validation and beyond: Next generation sequencing of forensic casework samples including challenging tissue samples from altered human corpses using the MiSeq FGx system

Senst¹,

Caliebe²,

Scheurer³

et al. 2022

Journal of Forensic Sciences

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“…Forensic specialists are challenged to work on human remains subjected to various environmental factors, including extremely high temperatures. The challenge is a driving force to search for optimal biological materials for post-mortem genetic analyses [12]. Apart from searching for "the best tissue," they also attempt to find the optimal location for collecting material during an autopsy [12] and optimal isolation methods for burnt biological material [13].…”

Section: Introductionmentioning

confidence: 99%

“…The challenge is a driving force to search for optimal biological materials for post-mortem genetic analyses [12]. Apart from searching for "the best tissue," they also attempt to find the optimal location for collecting material during an autopsy [12] and optimal isolation methods for burnt biological material [13]. In the case of burnt corpses, hard tissue is often the only material available for examination.…”

Section: Introductionmentioning

confidence: 99%

The intervertebral discs’ fibrocartilage as a DNA source for genetic identification in severely charred cadavers

Tomsia

Droździok

Banaszek

et al. 2022

Forensic Sci Med Pathol

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Identifying charred human remains poses a challenge to forensic laboratories. High temperature completely incinerates the superficial tissues and partially destroys bones, forcing the forensics to seek an alternative, for bones and teeth, forensic material that should quickly and cheaply deliver DNA of sufficient quantity and quality. We sought, other than rib cartilage, types of cartilages that could serve as a DNA source. DNA was isolated from the fibrous cartilage of a fibrous ring of intervertebral L1-L2 discs sampled from charred cadavers or charred body fragments: 5 victims of car fires, 1 victim of combustion during a residential house gas explosion, and 3 victims of nitroglycerin explosion. DNA was isolated by the column method. DNA quality and concentration were assessed by RT-PCR and multiplex PCR for 23 autosomal and 17 Y chromosome STR loci. STR polymorphism results obtained by capillary electrophoresis served for likelihood ratio (LR) calculations. DNA concentration in relation to the cadaver’s age and post-mortem interval (PMI) were analyzed. All samples (n = 9) yielded good-quality DNA in quantities (0.57–17.51 ng/µL for T. Large autosomal sequence) suitable for STR-based amplification. The isolated DNA characterized a low degradation index (0.80–1.99), and we were able to obtain complete genetic profiles. In each of the nine cases, the genotyping results allowed identifying the victims based on comparative material from the immediate family. The results demonstrate the usefulness of human intervertebral disc fibrocartilage as an alternative DNA source for the genetic identification of charred bodies or charred torso fragments.

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Analysis of the sequencing quality of next-generation sequencing for the entire mitochondrial genome in decomposed human samples

Lee,

Kim,

Lee

et al. 2024

Int J Legal Med

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Which tissue to take? A retrospective study of the identification success of altered human remains

Cited by 7 publications

References 22 publications

Validation and beyond: Next generation sequencing of forensic casework samples including challenging tissue samples from altered human corpses using the MiSeq FGx system

Validation and beyond: Next generation sequencing of forensic casework samples including challenging tissue samples from altered human corpses using the MiSeq FGx system

The intervertebral discs’ fibrocartilage as a DNA source for genetic identification in severely charred cadavers

Analysis of the sequencing quality of next-generation sequencing for the entire mitochondrial genome in decomposed human samples

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