Whose blood is it? Application of DEPArray™ technology for the identification of individual/s who contributed blood to a mixed stain

Anslinger, Katja; Bayer, Birgit

doi:10.1007/s00414-018-1912-7

Cited by 21 publications

(22 citation statements)

References 14 publications

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“…In particular, whole genome amplification of single CTCs and WBCs was carried out to confirm the absence of WBC contamination by DNA finger printing . Besides, similar results have been published in other application domains, where direct DNA fingerprinting of individual and pools of cells collected by DEPArray™ from forensic evidence has shown achievement of 100% purity .…”

Section: Accuracy Of Deparray™supporting

confidence: 54%

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

2018

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Circulating tumor cells (CTCs) are rare cells shed into the bloodstream by invasive tumors and their analysis offers a promising noninvasive tool to predict and monitor therapeutic responses. CTCs can be isolated from patient blood and their characterization at single‐cell level can inform on the genomic landscape of a tumor. All CTC enrichment methods bear a burden of contaminating normal cells, which mandate a further step of purification to enable reliable downstream genetic analysis. Here, we describe the DEPArray™ technology, a microchip‐based digital sorter, which combines precise microfluidic and microelectronic enabling precise, image‐based isolation of single CTCs, which can then be analyzed by Next Generation Sequencing (NGS) methods. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

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Section: Accuracy Of Deparray™supporting

confidence: 54%

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

2018

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“…10. and highly probative log (LR)s were recovered by DSCS from every donor (range [13][14][15][16][17][18][19][20][21][22][23][24]. No standard bulk mixture analysis results were available for comparison with this mixture due to computational limitations.…”

Section: Dscs Applied To Complex Mixturesmentioning

confidence: 99%

“…A potential solution to overcoming the issue of genotype information loss is to separate individual cells or a small subset of cells from the original 'bulk' mixture prior to analysis [10,[14][15][16][17][18][19][20][21]. Doing so may result in single source DNA profiles for all contributors as well as new 'mini mixtures' with less complexity compared to the original bulk mixture due to a decrease in the NOC [10].…”

Section: Introductionmentioning

confidence: 99%

Probabilistic Genotyping of Single Cell Replicates from Complex DNA Mixtures Recovers Higher Contributor LRs than Standard Analysis

Huffman

Hanson

Ballantyne

2021

Preprint

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DNA mixtures are a common source of crime scene evidence and are often one of the more difficult sources of biological evidence to interpret. With the implementation of probabilistic genotyping (PG), mixture analysis has been revolutionized allowing previously unresolvable mixed profiles to be analyzed and probative genotype information from contributors to be recovered. However, due to allele overlap, artifacts, or low-level minor contributors, genotype information loss inevitably occurs. In order to reduce the potential loss of significant DNA information from donors in complex mixtures, an alternative approach is to physically separate individual cells from mixtures prior to performing DNA typing thus obtaining single source profiles from contributors. In the present work, a simplified micromanipulation technique combined with enhanced single-cell DNA typing was used to collect one or few cells, referred to as direct single-cell subsampling (DSCS). Using this approach, single and 2-cell subsamples were collected from 2-6 person mixtures. Single-cell subsamples resulted in single source DNA profiles while the 2-cell subsamples returned either single source DNA profiles or new mini-mixtures that are less complex than the original mixture due to the presence of fewer contributors. PG (STRmix™) was implemented, after appropriate validation, to analyze the original bulk mixtures, single source cell subsamples, and the 2-cell mini mixture subsamples from the original 2-6-person mixtures. PG further allowed replicate analysis to be employed which, in many instances, resulted in a significant gain of genotype information such that the returned donor likelihood ratios (LRs) were comparable to that seen in their single source reference profiles (i.e., the reciprocal of their random match probabilities). In every mixture, the DSCS approach gave improved results for each donor compared to standard bulk mixture analysis. With the 5- and 6- person complex mixtures, DSCS recovered highly probative LRs (> 1020) from donors that had returned non-probative LRs (<103) by standard methods.

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“…A previous report has shown that isolation of multiple groups of cells from mixtures results in sub-populations with differing constituent weight ratios [ 6 ] that, in combination, can recover more DNA profile information, while others have demonstrated the potential to obtain highly probative single source samples from mixtures following analysis of recovered single or multiple cell subsets [ 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 ]. One of these latter physical cell recovery methods (direct single cell subsampling, DSCS) allows for multiple physical subsamplings of individual cells (or cell subsets) from complex cellular DNA mixtures using a stereomicroscope and tungsten needle.…”

Section: Introductionmentioning

confidence: 99%

Cell Subsampling Recovers Probative DNA Profile Information from Unresolvable/Undetectable Minor Donors in Mixtures

Huffman

Hanson

Ballantyne

2022

Genes

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When a minor DNA component to a binary mixture is present at a weight ratio of approximately 1:50 or less, the presence of this minor donor is undetectable (or barely detectable) by standard mixture deconvolution approaches. In an attempt to retrieve probative minor donor DNA profile information, multiple quintuple cell subsamples were collected from a 1:50 DNA mixture using direct single cell subsampling (DSCS) paired with probabilistic genotyping (PG), the latter validated for use with single or few cells. DSCS employs a simplified micromanipulation technique paired with an enhanced DNA profiling approach, involving direct cell lysis and a sensitive PCR process, to genotype individual cells. Multiple five-cell subsamples were used to interrogate sufficient cells from the mixture such that some of the created 5-cell “mini-mixture” subsamples contained a cell from the minor donor. The latter mini-mixture subsamples, which now comprised weight ratios of 1:4 as opposed to the bulk mixture 1:50, were analyzed with the PG systems STRmixTM and EuroForMix resulting in a significant probative gain of information, (LR ≅ 1011, compared to standard bulk mixture PG methods, LR ≅ 101–102).

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Whose blood is it? Application of DEPArray™ technology for the identification of individual/s who contributed blood to a mixed stain

Cited by 21 publications

References 14 publications

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

Probabilistic Genotyping of Single Cell Replicates from Complex DNA Mixtures Recovers Higher Contributor LRs than Standard Analysis

Cell Subsampling Recovers Probative DNA Profile Information from Unresolvable/Undetectable Minor Donors in Mixtures

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