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<p> A&#xa0;lateral flow biosensor was developed&#xa0;based on newly established primer extension nucleic acid isothermal amplification (PEIA) and M13mp18 single-stranded DNA (ssDNA) to visually identify the H1N1 virus. The PEIA process was primer based on a template to continuously extend multiple repeat sequences, using DNA polymerase, to form long ssDNAs. M13mp18 ssDNA was then coupled to multiple long ssDNAs from PEIA and functionalized in a signal tag DNA nanostructure (STDN) for the lateral flow biosensor. In the presence of target H1N1-RNA, the sandwich structure capture probe/H1N1-RNA/STDN is fabricated caught in the lateral flow biosensor test zone, and appears red. Our biosensor successfully identified H1N1 virus RNA over a broad range of concentrations (10–10<sup>6</sup>&#xa0;pM) with a minimum visual detection limit of 10&#xa0;pM. In conclusion, our biosensor may be applied to point-of-care diagnostics in low-resource environments with non-specialist staff.</p> Graphical Abstract <p></p>

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Lateral flow biosensor development for the visual identification of H1N1 virus based on primer extension nucleic acid isothermal amplification and M13mp18 single-stranded DNA

  • 期刊论文

  • 发表日期：

    2025年2月18日

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  • Lingling Wang,
  • Shengjun Bu,
  • Heyuan Meng,
  • Xiaoying Zhang,
  • Hongyu Zhou,
  • Chuanna Xu,
  • Zhuo Hao,
  • Yuchun Su,
  • Jiayu Wan

摘要

A lateral flow biosensor was developed based on newly established primer extension nucleic acid isothermal amplification (PEIA) and M13mp18 single-stranded DNA (ssDNA) to visually identify the H1N1 virus. The PEIA process was primer based on a template to continuously extend multiple repeat sequences, using DNA polymerase, to form long ssDNAs. M13mp18 ssDNA was then coupled to multiple long ssDNAs from PEIA and functionalized in a signal tag DNA nanostructure (STDN) for the lateral flow biosensor. In the presence of target H1N1-RNA, the sandwich structure capture probe/H1N1-RNA/STDN is fabricated caught in the lateral flow biosensor test zone, and appears red. Our biosensor successfully identified H1N1 virus RNA over a broad range of concentrations (10–106 pM) with a minimum visual detection limit of 10 pM. In conclusion, our biosensor may be applied to point-of-care diagnostics in low-resource environments with non-specialist staff.

Graphical Abstract