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

TL;DR

A new method using a heat-resistant enzyme and a nuclease to detect bacteria in food contamination is developed, offering a flexible and sensitive detection tool.

Contribution

A primer design using deoxy inosine base with endonuclease V to activate PfAgo for universal nucleic acid detection is introduced.

Findings

The detection limit of the method is 0.04 ng/μL DNA.

The method successfully detects food contaminated bacteria.

The approach is a universal tool for detecting nucleic acid-containing organisms like pathogens and viruses.

Abstract

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.

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