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<p>Despite extraordinary efforts to develop rapid molecular diagnostics for bacteremia, accurate and timely detection remains a challenge. While most molecular technologies are designed to detect bacteria circulating in the bloodstream, few technologies that detect bacteria engulfed by circulating neutrophils have been developed. Here, we describe a single-cell digital PCR assay to quantify circulating neutrophils harboring phagocytosed bacteria. We encapsulated a single neutrophil within a single droplet of a PCR reaction mix, and then performed in situ PCR to detect intra-neutrophil bacterial DNA. In this way, we successfully quantified neutrophils containing <i>Staphylococcus aureus</i> DNA, as well as human genes, at single-cell resolution. This technology, termed droplet digital cell PCR, enables rapid and accurate identification of bacteria directly from blood, thereby improving diagnosis and management of bacteremia.</p>

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Quantifying neutrophil phagocytes by droplet digital cell PCR for diagnosis of bacteremia

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月20日

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  • Naoki Uno,
  • Toshifumi Imagawa,
  • Kenji Ota,
  • Norihito Kaku,
  • Katsunori Yanagihara,
  • Yuki Furuse

摘要

Despite extraordinary efforts to develop rapid molecular diagnostics for bacteremia, accurate and timely detection remains a challenge. While most molecular technologies are designed to detect bacteria circulating in the bloodstream, few technologies that detect bacteria engulfed by circulating neutrophils have been developed. Here, we describe a single-cell digital PCR assay to quantify circulating neutrophils harboring phagocytosed bacteria. We encapsulated a single neutrophil within a single droplet of a PCR reaction mix, and then performed in situ PCR to detect intra-neutrophil bacterial DNA. In this way, we successfully quantified neutrophils containing Staphylococcus aureus DNA, as well as human genes, at single-cell resolution. This technology, termed droplet digital cell PCR, enables rapid and accurate identification of bacteria directly from blood, thereby improving diagnosis and management of bacteremia.