A Rapid and Visual Method for Nucleic Acid Detection of Escherichia coli O157:H7 Based on CRISPR/Cas12a-PMNT

被引：12
作者：
Jiang, Wei [1 ,2 ,3 ]
He, Chuan [1 ,2 ,3 ]
Bai, Lan [1 ,2 ,3 ]
Chen, Yifan [1 ,2 ,3 ]
Jia, Junwei [1 ,3 ]
Pan, Aihu [2 ,3 ]
Lv, Beibei [1 ,2 ,3 ]
Tang, Xueming [1 ,2 ,3 ,4 ]
Wu, Xiao [1 ,2 ,3 ]
机构：
[1] Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China
[2] Shanghai Acad Agr Sci, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
[3] Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspect & Supervis Ctr S, Shanghai 201106, Peoples R China
[4] Shanghai Jiao Tong Univ, Sch Agr & Biol, Shanghai 200240, Peoples R China
来源：
FOODS | 2023年 / 12卷 / 02期
关键词：
foodborne pathogenic bacteria; Escherichia coli O157; H7; nucleic acid detection; POCT; visual method; CRISPR-Cas12a; PMNT; STAPHYLOCOCCUS-AUREUS;
D O I：
10.3390/foods12020236
中图分类号：
TS2 [食品工业];
学科分类号：
0832 ;
摘要：
Rapid, accurate and visual point-of-care testing (POCT) methods for pathogenic bacteria detection are essential for avoiding foodborne diseases caused by pathogens or their toxins. In this study, we proposed a rapid and visual detection method that we named "Cas12aVIP". By combining recombinase polymerase amplification (RPA), a CRISPR/Cas12a system and a cationic-conjugated polythiophene derivative (poly[3-(3 '-N,N,N-triethylamino-1 '-propyloxy)-4-methyl-2,5-thiophene hydrochloride] (PMNT) mixed with single-stranded DNA (ssDNA)), the solution turned red in the absence of the target DNA based on conformational modifications of the conjugated backbone of PMNT, whereas it displayed yellow, thus realizing the colorimetric detection of DNA. The Cas12aVIP method yielded high specificity and no interference from other nontargeted bacteria. The detection was accomplished in 40 min and the signal could be observed by the naked eye under natural light, presenting great potential for a variety of rapid nucleic acid detection applications without requiring technical expertise or ancillary equipment.
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页数：7
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