文献类型： 外文期刊
作者： Wang, Liu 1 ; Bai, Linlin 1 ; Wang, Hongmei 1 ; He, Kaiyu 1 ; Wang, Rui 2 ; Wang, Qiang 1 ; Zhang, Fang 3 ; Xu, Xiahong 1 ;
作者机构： 1.Zhejiang Acad Agr Sci, Inst Agroprod Safety & Nutr, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou 310021, Peoples R China
2.Fudan Univ, Human Phenome Inst, State Key Lab Genet Engn, Shanghai 200438, Peoples R China
3.Fuzhou Univ, Coll Biol Sci & Engn, Fuzhou 350108, Peoples R China
4.Minist Agr & Rural Affairs, Key Lab Traceabil Agr Genet Modified Organisms, Hangzhou 310021, Peoples R China
5.Minist Agr & Rural Affairs, Key Lab Informat Traceabil Agr Prod, Hangzhou 310021, Peoples R China
关键词： Salmonella; CRISPR/Cas12a; Graphene oxide; Visual detection; Fluorescent recovery
期刊名称：MICROCHEMICAL JOURNAL （ 影响因子：4.8； 五年影响因子：4.5 ）
ISSN： 0026-265X
年卷期： 2023 年 191 卷
页码：
收录情况： SCI
摘要： Rapid and accurate detection of Salmonella is extremely important to ensure food safety. Combining nucleic acid amplification with CRISPR/Cas12a can realize sensitive and specific detection of foodborne pathogens. However, present CRISPR/Cas-based methods mainly depend on fluorophore and quencher dual-labeled ssDNA as the reporter, which brings in high cost and steric hindrance. Herein, a new signal output format has been developed. The method takes advantage of graphene oxide adsorbing FAM-ssDNA and quenching the fluorescent emission while CRISPR/Cas12a recovering the fluorescent emission by trans-cleaving the FAM-ssDNA. Factors affecting the performance of the sensor have been optimized, and it is shown that the oligonucleotide of 12 nt with the fluorophore modified at the 3 ' end displays the highest signal to noise ratio. The signal output method has been combined with recombinase polymerase amplification for detection of Salmonella and presents consistent specificity and sensitivity (5 x 101 copies) as real-time PCR. By replacing the dual-labeled ssDNA reporter with FAM-ssDNA and GO, this signal output method saves similar to 65% cost. Notably, this method does not require a real-time thermal cycler or any other sophisticated instrument. Besides pathogens, this easy and convenient method can also be extended to detect other targets.
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