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<p><i>Pyrococcus furiosus</i> Argonaute (<i>Pf</i>Ago) is a novel programmable nuclease that has been used in nucleic acid detection due to its excellent performance. Traditional <i>Pf</i>Ago based detection methods relies on the input of exogenous guide DNA (gDNA), which restricted its flexibility and universality in application. Here, we designed primers with deoxy inosine base which can be recognized and cleaved by endonuclease V, following turn into gDNA to activate <i>Pf</i>Ago for target gene detection. Therefore, an Endo <Emphasis Type="Underline">V</Emphasis> activated <Emphasis Type="ItalicUnderline">P</Emphasis><i>f</i>Ago based <Emphasis Type="Underline">n</Emphasis>ucleic acid detection (VPN) method was developed. The detection limit of this method was 0.04 ng/μL DNA. Moreover, the method was successfully applied to the detection of food contaminated bacteria. This approach provided a universal and power tool for the detection of nucleic acid-containing organisms such as pathogens, viruses, and tumor cells.</p><p></p>

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Endonuclease V activated Pyrococcus furiosus Argonaute for the detection of food contaminated bacteria

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月31日

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  • Yiheng Shi,
  • Pei Gao,
  • Di Wu,
  • Yongning Wu,
  • Guoliang Li

摘要

Pyrococcus furiosus Argonaute (PfAgo) is a novel programmable nuclease that has been used in nucleic acid detection due to its excellent performance. Traditional PfAgo based detection methods relies on the input of exogenous guide DNA (gDNA), which restricted its flexibility and universality in application. Here, we designed primers with deoxy inosine base which can be recognized and cleaved by endonuclease V, following turn into gDNA to activate PfAgo for target gene detection. Therefore, an Endo V activated PfAgo based nucleic acid detection (VPN) method was developed. The detection limit of this method was 0.04 ng/μL DNA. Moreover, the method was successfully applied to the detection of food contaminated bacteria. This approach provided a universal and power tool for the detection of nucleic acid-containing organisms such as pathogens, viruses, and tumor cells.