Details

Autor(en) / Beteiligte

• Shi, Yiheng
• Tan, Zishan
• Wu, Di
• Wu, Yongning
• Li, Guoliang

Titel

Pyrococcus furiosus argonaute based Alicyclobacillus acidoterrestrsis detection in fruit juice

Ist Teil von

• Food microbiology, 2024-06, Vol.120, p.104475-104475, Article 104475

Ort / Verlag

England: Elsevier Ltd

Erscheinungsjahr

2024

Quelle

Access via ScienceDirect (Elsevier)

Beschreibungen/Notizen

• Alicyclobacillus acidoterrestris is the major threat to fruit juice for its off-odor producing characteristic. In this study, Pyrococcus furiosus Argonaute (PfAgo), a novel endonuclease with precise DNA cleavage activity, was used for A. acidoterrestrisdetection, termed as PAD. The partially amplified 16 S rRNA gene of A. acidoterrestris can be cleaved by PfAgo activated by a short 5′-phosphorylated single strand DNA, producing a new guide DNA (gDNA). Then, PfAgo was activated by the new gDNA to cut a molecular beacon (MB) with fluorophore-quencher reporter, resulting in the recovery of fluorescence. The fluorescent intensity is positively related with the concentration of A. acidoterrestris. The PAD assay showed excellent specificity and sensitivity as low as 101 CFU/mL, which can be a powerful tool for on-site detection of A. acidoterrestris in fruit juice industry in the future, reducing the economic loss. •A novel nucleic acid detection method based on argonaute for A. acidoterrestris was developed.•The assay showed excellent specificity and sensitivity with a limit of detection of 101 CFU/mL.•The assay exhibited good performance in real sample analysis.