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<p>Mixed biological stains are commonly encountered biological samples in forensic investigation. They are often challenging particularly when containing contributors of the same tissue type, such as mixed bloodstains. Here, we described a novel genetic marker for the separation of two-individual mixed bloodstain: the human neutrophil antigen (HNA), specifically, the HNA-1 system. HNA-1a and HNA-1b exhibit the high polymorphism across diverse populations. Potential for practical applicability of these markers in mixed blood separation was evaluated through fluorescence-activated cell sorting (FACS) and laser capture microdissection (LCM) techniques. The results found that flow cytometry based on HNA-1a and HNA-1b fluorescent antibody successfully separated minor contributors from mixed samples at ratios as low as 1:16 and 1:32, respectively. Furthermore, LCM enabled complete single-source STR profiling from the fewer cells, indicating promising application in the separation of mixed bloodstains. Collectively, this study provides a new usable genetic marker, which may significantly enhance the effectiveness of antigen-based separation strategies for the identification of mixed blood samples.</p>

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Separation and identification of two-individual mixed bloodstains using human neutrophil antigen-1 antibodies

  • 期刊论文

  • 发表日期：

    2026年1月27日

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  • Xi Xia,
  • Ji-long Zheng,
  • Si-wen Wang,
  • Zhenze Liu,
  • Zhou Li,
  • Dan-yang Wang,
  • Xiao-na Li,
  • Xu Wu,
  • Jun Yao

摘要

Mixed biological stains are commonly encountered biological samples in forensic investigation. They are often challenging particularly when containing contributors of the same tissue type, such as mixed bloodstains. Here, we described a novel genetic marker for the separation of two-individual mixed bloodstain: the human neutrophil antigen (HNA), specifically, the HNA-1 system. HNA-1a and HNA-1b exhibit the high polymorphism across diverse populations. Potential for practical applicability of these markers in mixed blood separation was evaluated through fluorescence-activated cell sorting (FACS) and laser capture microdissection (LCM) techniques. The results found that flow cytometry based on HNA-1a and HNA-1b fluorescent antibody successfully separated minor contributors from mixed samples at ratios as low as 1:16 and 1:32, respectively. Furthermore, LCM enabled complete single-source STR profiling from the fewer cells, indicating promising application in the separation of mixed bloodstains. Collectively, this study provides a new usable genetic marker, which may significantly enhance the effectiveness of antigen-based separation strategies for the identification of mixed blood samples.