Ultrasensitive detection and rapid identification of multiple foodborne pathogens with the naked eyes

• Published in: Biosensors & bioelectronics Vol. 71; pp. 186 - 193
• Main Authors: Zhang, Heng, Zhang, Yali, Lin, Yankui, Liang, Tongwen, Chen, Zhihua, Li, Jinfeng, Yue, Zhenfeng, Lv, Jingzhang, Jiang, Qing, Yi, Changqing
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 15.09.2015
• Subjects: Amplification; Arrays; Biosensing Techniques - instrumentation; Colorimetric; Colorimetry - economics; Colorimetry - instrumentation; Deoxyribonucleic acid; DNA microarray; DNA, Bacterial - analysis; DNA, Bacterial - genetics; Escherichia coli; Escherichia coli O157 - genetics; Escherichia coli O157 - isolation & purification; Eyes; Food Contamination - analysis; Food Microbiology - economics; Food Microbiology - instrumentation; Foodborne pathogens; Genes; Humans; Limit of Detection; Listeria monocytogenes; Listeria monocytogenes - genetics; Listeria monocytogenes - isolation & purification; Multiple cycle signal amplification; Naked eye detection; Oligonucleotide Array Sequence Analysis - economics; Oligonucleotide Array Sequence Analysis - instrumentation; Panels; Pathogens; Polymerase Chain Reaction - economics; Polymerase Chain Reaction - instrumentation; Reproduction; Salmonella enterica; Salmonella enterica - genetics; Salmonella enterica - isolation & purification; Time Factors; DNA microarray; Foodborne pathogens; Naked eye detection; Multiple cycle signal amplification; Colorimetric
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2015.04.034

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. •The naked eye detection of multiple foodborne pathogens with ultrahigh sensitivity is realized.•Colorimetric detection combined with PCR offers advantages of easy manipulation, high efficiency and low cost.•The whole detection involved a hybridization procedure and an additional multiple cycle signal amplification procedure.•The detectable signal is amplified by the repeated staining of biotinated DNA duplexes with AV-HRP and biotinated anti-HRP antibody.•This simple colorimetric assay shows excellent anti-interference capability and good stability.