DNA extraction for short tandem repeat typing from mixed samples using anti-human leukocyte CD45 and ABO blood group antibodies

Yano, Shizue; Honda, Katsuya; Kaminiwa, Junko; Nishi, Takeki; Iwabuchi, Yayoi; Sugano, Yukiko; Kurosu, Akira; Suzuki, Yasuhito

doi:10.1016/j.fsigen.2014.01.001

Cited by 12 publications

(9 citation statements)

References 16 publications

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“…These include the separation of different cell types by laser capture microdissection (LCM) or fluorescence-activated cell sorting (FACS) [6,18], as well as the time-shift release of the DNA from the different cellular components of a mixture, such as the differential extraction method for sperm-epithelial cell mixtures [10]. For the splitting of mixed stains consisting exclusively of cells of the same cell type, some other methods may be helpful; for example, the use of anti-human AB0 and CD45 antibody-coated microbeads combined with centrifugal separation for the isolation of white blood cells originating from donors with different blood groups [19]. As a further approach, the sex-specific labelling of cells combined with laser microdissection is used for the isolation of cells from a male and a female contributor [1,2].…”

Section: Mixture Deconvolutionmentioning

confidence: 99%

Deconvolution of blood-blood mixtures using DEPArrayTM separated single cell STR profiling

2018

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Background. So far STR (Short Tandem Repeat) profiling of single cells has played hardly any role in forensic molecular-biological casework; however, the application of this technique could open up new possibilities for deducing the unambiguous DNA profile of each single contributor involved in a mixture of two or more components. Objective. For this purpose DEPArray TM technology was used to isolate single white blood cells from mixed stains containing blood from two or three different contributors. The STR profiling was carried out on each single cell. Material and methods. In order to standardize the interpretation of the findings, which, like all low template (LT) DNA analyses, shows characteristic artifacts, leukocytes from a fresh blood sample were examined in advance and

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Section: Mixture Deconvolutionmentioning

confidence: 99%

Deconvolution of blood-blood mixtures using DEPArrayTM separated single cell STR profiling

2018

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“…The main approach to examine the concept of mixture separation so far is the use of laser capture microdissection [12]; although, to our knowledge, this has not been applied to separate blood from epithelial cells. Recently, the use of anti-CD45 labelled magnetic beads and ABO antibodies were trialled to separate leukocytes from mixed bloodstains [13], with results indicating that further research may lead to cleaner profiles from mixed stains containing blood.…”

Section: Introductionmentioning

confidence: 99%

“…As a wide variety of antibody markers are commercially available, choices which reflect cell types of interest, e.g. anti-CD45 (a white blood cell surface antigen [17] as used by Yano et al [13]) and anti-CD227 (a mucosal epithelial cell surface antigen [18]), can be readily made.…”

Section: Introductionmentioning

confidence: 99%

FACS separation of non-compromised forensically relevant biological mixtures

Verdon

Mitchell

Chen

et al. 2015

Forensic Science International: Genetics

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“…In recent studies, flow cytometry has also been successfully applied to identify and separate sperm cells from vaginal epithelial cell mixtures in forensic science922. However, only scarce data has been published applying this approach to mixed samples from two or more donors with the same gender111223. We, for the first time, aimed to identify single donor’s genotype from mixed sperm cells involving plural contributors by FACS.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence- and magnetic-activated cell sorting strategies to separate spermatozoa involving plural contributors from biological mixtures for human identification

Xie

Chen

et al. 2016

Sci Rep

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No effective method has been developed to distinguish sperm cells originating from different men in multi-suspect sexual assault cases. Here we combined MACS and FACS to isolate single donor sperm cells from forensic mixture samples including female vaginal epithelial cells and sperm cells from multiple contributors. Sperms from vaginal swab were isolated by MACS using FITC-conjugated A kinase anchor protein 3 (AKAP3) antibody; target individual sperm cells involving two or three donors were separated by FACS using FITC-labeled blood group A/B antigen antibody. This procedure was further tested in two mock multi-suspect sexual assault samples and one practical casework sample. Our results showed that complete single donor STR profiles could be successfully obtained from sperm/epithelial cell and sperm mixtures from two contributors. For unbalanced sperm/epithelial cells and sperm cells mixtures, sensitivity results revealed that target cells could be detected at as low as 1:32 and 1:8 mixed ratios, respectively. Although highly relies on cell number and blood types or secretor status of the individuals, this procedure would still be useful tools for forensic DNA analysis of multi-suspect sexual assault cases by the combined use of FACS and MACS based on sperm-specific AKAP3 antigen and human blood type antigen.

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DNA extraction for short tandem repeat typing from mixed samples using anti-human leukocyte CD45 and ABO blood group antibodies

Cited by 12 publications

References 16 publications

Deconvolution of blood-blood mixtures using DEPArrayTM separated single cell STR profiling

Deconvolution of blood-blood mixtures using DEPArrayTM separated single cell STR profiling

FACS separation of non-compromised forensically relevant biological mixtures

Fluorescence- and magnetic-activated cell sorting strategies to separate spermatozoa involving plural contributors from biological mixtures for human identification

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