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<p><i>Haemophilus influenzae</i>, particularly type b (Hib), represents a significant opportunistic pathogen that poses a substantial threat to the health and safety of vulnerable populations, including the elderly and children. To facilitate early detection and timely intervention, there is an urgent need for a simple, portable, and efficient molecular diagnostic approach. In this study, a duplex recombinase polymerase amplification assay with a lateral flow strip was developed to target the outer membrane protein P6 (<i>omp</i>6) of <i>H. influenzae</i>, and the capsule encoding gene <i>bex</i>A of capsulated <i>H. influenzae.</i> The assay is performed at 37℃ for 25&#xa0;min, with a limit of detection of 10<sup>1</sup> CFU, comparable to that of dual PCR. The detection accuracy for clinical samples was consistent with the traditional culture and dual PCR methods. The duplex RPA assay does not require a laboratory facility and may be useful for genotyping of <i>H. influenzae.</i></p>

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Simultaneous visual diagnosis of capsulated and non-capsulated Haemophilus influenzae using a duplex RPA method

  • 期刊论文

  • 发表日期：

    2025年11月18日

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  • Lei Wang,
  • Jun Zhu,
  • Wenxue Liang,
  • Juan Huo,
  • Ting Zhang,
  • Yan Wang

摘要

Haemophilus influenzae, particularly type b (Hib), represents a significant opportunistic pathogen that poses a substantial threat to the health and safety of vulnerable populations, including the elderly and children. To facilitate early detection and timely intervention, there is an urgent need for a simple, portable, and efficient molecular diagnostic approach. In this study, a duplex recombinase polymerase amplification assay with a lateral flow strip was developed to target the outer membrane protein P6 (omp6) of H. influenzae, and the capsule encoding gene bexA of capsulated H. influenzae. The assay is performed at 37℃ for 25 min, with a limit of detection of 101 CFU, comparable to that of dual PCR. The detection accuracy for clinical samples was consistent with the traditional culture and dual PCR methods. The duplex RPA assay does not require a laboratory facility and may be useful for genotyping of H. influenzae.