Endoprotein-activating DNAzyme assay for nucleic acid extraction- and amplification-free detection of viable pathogenic bacteria

被引:3
作者
Wu, Zixiang [1 ]
Chen, Yanbai [1 ]
Wei, Junlun [3 ]
Deng, Yi [1 ]
Deng, Ruijie [2 ]
Yang, Hao [1 ,2 ]
机构
[1] Sichuan Univ, Sch Chem Engn, Chengdu 610065, Peoples R China
[2] Sichuan Univ, Coll Biomass Sci & Engn, Chengdu 610065, Peoples R China
[3] Sichuan Univ, West China Hosp, Ctr Diabet & Metab Res, Dept Endocrinol & Metab, Chengdu 610041, Peoples R China
关键词
DNAzyme; Amplification-free; Viable pathogenic bacteria; RNase H2; Fluorescent assay; SALMONELLA;
D O I
10.1016/j.bios.2024.116715
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Pathogenic bacteria in food or environment, can pose threats to public health, highlighting the requirement of tools for rapid and accurate detection of viable pathogenic bacteria. Herein, we report a sequential endoprotein RNase H2-activating DNAzyme assay (termed epDNAzyme) that enables nucleic acid extraction- and amplification-free detection of viable Salmonella enterica (S. S. enterica). ). The direct detection allows for a rapid detection of viable S. enterica within 25 min. Besides, the assay, based on sequential reporting strategy, circumvents internal modifications in the DNAzyme's active domain and improve its catalytic activity. The multiple-turnover DNAzyme cutting and the enhanced catalytic activity of DNAzyme render the epDNAzyme assay to be highly sensitive, and enables the detection of 190 CFU/mL and 0.1% viable S. enterica. . The assay has been utilized to detect S. enterica contamination in food and clinical samples, indicating its potential as a promising tool for monitoring pathogen-associated biosafety.
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页数:7
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