title: Ultrasensitive detection and rapid identification of multiple foodborne pathogens with the naked eyes.
abstract: In this study, a novel approach for ultrasensitive detection and rapid high-throughput identification of a panel of common foodborne pathogens with the naked eyes is presented. As a proof-of-concept application, a multiple pathogen analysis array is fabricated through immobilizing three specific polyT-capture probes which can respectively recognize rfbE gene (Escherichia coli O157:H7), invA gene (Salmonella enterica), inlA gene (Listeria monocytogenes) on the plastic substrates. PCR has been developed for amplification and labeling target genes of rfbE, invA, inlA with biotin. The biotinated target DNA is then captured onto the surface of plastic strips through specific DNA hybridization. The succeeding staining of biotinated DNA duplexes with avidin-horseradish peroxidise (AV-HRP) and biotinated anti-HRP antibody greatly amplifies the detectable signal through the multiple cycle signal amplification strategy, and thus realizing ultrasensitive and specific detection of the above three pathogens in food samples with the naked eyes. Results showed approximately 5 copies target pathogenic DNA could be detected with the naked eyes. This simple but very efficient colorimetric assay also show excellent anti-interference capability and good stability, and can be readily applied to point-of-care diagnosis.
authors.affiliation: Key Laboratory of Sensing Technology and Biomedical Instruments (Guangdong Province), School of Engineering, Sun Yat-Sen University, Guangzhou, China; Shenzhen Key Research Laboratory of Detection Technology R&D on Food Safety, Technical Centre for Food Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, China; Shenzhen Academy of Inspection and Quarantine, Shenzhen, China.
authors.forename: Heng
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Sensing Technology and Biomedical Instruments (Guangdong Province), School of Engineering, Sun Yat-Sen University, Guangzhou, China.
authors.forename: Yali
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Shenzhen Key Research Laboratory of Detection Technology R&D on Food Safety, Technical Centre for Food Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, China; Shenzhen Academy of Inspection and Quarantine, Shenzhen, China.
authors.forename: Yankui
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Shenzhen Key Research Laboratory of Detection Technology R&D on Food Safety, Technical Centre for Food Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, China; Shenzhen Academy of Inspection and Quarantine, Shenzhen, China.
authors.forename: Tongwen
authors.initials: T
authors.lastname: Liang
authors.affiliation: Key Laboratory of Sensing Technology and Biomedical Instruments (Guangdong Province), School of Engineering, Sun Yat-Sen University, Guangzhou, China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Shenzhen Key Research Laboratory of Detection Technology R&D on Food Safety, Technical Centre for Food Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, China; Shenzhen Academy of Inspection and Quarantine, Shenzhen, China.
authors.forename: Zhenfeng
authors.initials: Z
authors.lastname: Yue
authors.affiliation: Shenzhen Key Research Laboratory of Detection Technology R&D on Food Safety, Technical Centre for Food Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, China; Shenzhen Academy of Inspection and Quarantine, Shenzhen, China.
authors.forename: Jingzhang
authors.initials: J
authors.lastname: Lv
authors.affiliation: Key Laboratory of Sensing Technology and Biomedical Instruments (Guangdong Province), School of Engineering, Sun Yat-Sen University, Guangzhou, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Sensing Technology and Biomedical Instruments (Guangdong Province), School of Engineering, Sun Yat-Sen University, Guangzhou, China. Electronic address: yichq@mail.sysu.edu.cn.
authors.forename: Changqing
authors.initials: C
authors.lastname: Yi
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation; D003124:Colorimetry / Q000191:economics / Q000295:instrumentation*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology / Q000191:economics / Q000295:instrumentation; D006801:Humans; D057230:Limit of Detection; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000295:instrumentation; D016133:Polymerase Chain Reaction / Q000191:economics / Q000295:instrumentation; D019779:Salmonella enterica / Q000235:genetics / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D004269:DNA, Bacterial
keywords: Colorimetric; DNA microarray; Foodborne pathogens; Multiple cycle signal amplification; Naked eye detection