Immuno-capture and in situ detection of Salmonella typhimurium on a novel microfluidic chip.

The new method presented in this article achieved the goal of capturing Salmonella typhimurium via immunoreaction and rapid in situ detection of the CdSe/ZnS quantum dots (QDs) labeled S. typhimurium by self-assembly light-emitting diode-induced fluorescence detection (LIF) microsystem on a specially designed multichannel microfluidic chip. CdSe/ZnS QDs were used as fluorescent markers improving detection sensitivity. The microfluidic chip developed in this study was composed of 12 sample channels, 3 mixing zones, and 6 immune reaction zones, which also acted as fluorescence detection zones. QDs-IgG-primary antibody complexes were generated by mixing CdSe/ZnS QDs conjugated secondary antibody (QDs-IgG) and S. typhimurium antibody (primary antibody) in mixing zones. Then, the complexes went into immune reaction zones to label previously captured S. typhimurium in the sandwich mode. The capture rate of S. typhimurium in each detection zone was up to 70%. The enriched QDs-labeled S. typhimurium was detected using a self-assembly LIF microsystem. A good linear relationship was obtained in the range from 3.7×10 to 3.7×10(5) cfu mL(-1) using the equation I=0.1739 log (C)-0.1889 with R(2)=0.9907, and the detection limit was down to 37 cfu mL(-1). The proposed method of online immunolabeling with QDs for in situ fluorescence detection on the designed multichannel microfluidic chip had been successfully used to detect S. typhimurium in pork sample, and it has shown potential advantages in practice.

[1]  Utkan Demirci,et al.  Portable microfluidic chip for detection of Escherichia coli in produce and blood , 2012, International journal of nanomedicine.

[2]  Jeong-Yeol Yoon,et al.  Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package. , 2013, Biosensors & bioelectronics.

[3]  D. Lim,et al.  A rapid and automated fiber optic-based biosensor assay for the detection of Salmonella in spent irrigation water used in the sprouting of sprout seeds. , 2004, Journal of food protection.

[4]  D. Pang,et al.  One-step sensitive detection of Salmonella typhimurium by coupling magnetic capture and fluorescence identification with functional nanospheres. , 2013, Analytical chemistry.

[5]  R. Levin,et al.  Efficacy of coating activated carbon with milk proteins to prevent binding of bacterial cells from foods for PCR detection. , 2013, Journal of microbiological methods.

[6]  Il-Hoon Cho,et al.  In-situ immuno-gold nanoparticle network ELISA biosensors for pathogen detection. , 2013, International journal of food microbiology.

[7]  Thierry Livache,et al.  On-chip microbial culture for the specific detection of very low levels of bacteria. , 2013, Lab on a chip.

[8]  Yuanying Chen,et al.  Rapid evolution of piRNA clusters in the Drosophila melanogaster ovary , 2023, bioRxiv.

[9]  J. Ji,et al.  Sensitive quantification and visual detection of bacteria using CdSe/ZnS@SiO2 nanoparticles as fluorescent probes , 2014 .

[10]  M. Prodromidis,et al.  Development of an impedimetric immunosensor based on electropolymerized polytyramine films for the direct detection of Salmonella typhimurium in pure cultures of type strains and inoculated real samples. , 2008, Analytica chimica acta.

[11]  Hye-Weon Yu,et al.  Electrochemical immunoassay using quantum dot/antibody probe for identification of cyanobacterial hepatotoxin microcystin-LR , 2009, Analytical and bioanalytical chemistry.

[12]  Alfredo Sanz-Medel,et al.  Quantum dot-based array for sensitive detection of Escherichia coli , 2011, Analytical and bioanalytical chemistry.

[13]  Hui Chen,et al.  Immunomagnetic separation and rapid detection of bacteria using bioluminescence and microfluidics. , 2009, Talanta.

[14]  Ihab Abdel-Hamid,et al.  Highly sensitive flow-injection immunoassay system for rapid detection of bacteria , 1999 .

[15]  Nuo Duan,et al.  A dual-color flow cytometry protocol for the simultaneous detection of Vibrio parahaemolyticus and Salmonella typhimurium using aptamer conjugated quantum dots as labels. , 2013, Analytica chimica acta.

[16]  S. Pillai,et al.  A microfluidic device for continuous capture and concentration of microorganisms from potable water. , 2007, Lab on a chip.

[17]  Guneet Singh Kohli,et al.  Highly sensitive detection of Salmonella typhi using surface aminated polycarbonate membrane enhanced-ELISA. , 2012, Biosensors & bioelectronics.

[18]  Chunyu Li,et al.  The dual role of deposited microbead plug (DMBP): a blood filter and a conjugate reagent carrier toward point-of-care microfluidic immunoassay. , 2012, Talanta.

[19]  Miguel Valcárcel,et al.  Colistin-functionalised CdSe/ZnS quantum dots as fluorescent probe for the rapid detection of Escherichia coli. , 2011, Biosensors & bioelectronics.

[20]  J. Jung,et al.  An integrated passive micromixer-magnetic separation-capillary electrophoresis microdevice for rapid and multiplex pathogen detection at the single-cell level. , 2011, Lab on a chip.

[21]  Jihea Moon,et al.  Detection of pathogenic Salmonella with nanobiosensors , 2013 .

[22]  D. R. Ess,et al.  An Automated Fiber-optic Biosensor Based Binding Inhibition Assay for the Detection of Listeria Monocytogenes , 2007 .

[23]  Frances S Ligler,et al.  Antimicrobial peptide-based array for Escherichia coli and Salmonella screening. , 2006, Analytica chimica acta.

[24]  Y. Lo,et al.  Highly sensitive and selective sensor based on silica-coated CdSe/ZnS nanoparticles for Cu2+ ion detection , 2012 .

[25]  Arun K. Bhunia,et al.  Detection of Low Levels of Listeria monocytogenes Cells by Using a Fiber-Optic Immunosensor , 2004, Applied and Environmental Microbiology.

[26]  Liju Yang,et al.  Dielectrophoresis assisted immuno-capture and detection of foodborne pathogenic bacteria in biochips. , 2009, Talanta.

[27]  Nobuyasu Yamaguchi,et al.  Rapid, Semiautomated Quantification of Bacterial Cells in Freshwater by Using a Microfluidic Device for On-Chip Staining and Counting , 2010, Applied and Environmental Microbiology.

[28]  Li Zhang,et al.  Rapid detection of pathogens using antibody-coated microbeads with bioluminescence in microfluidic chips , 2010, Biomedical microdevices.

[29]  Yongxin Song,et al.  Counting bacteria on a microfluidic chip. , 2010, Analytica chimica acta.

[30]  M. Yeh,et al.  Salmonella detection using 16S ribosomal DNA/RNA probe-gold nanoparticles and lateral flow immunoassay. , 2013, Food chemistry.

[31]  Miao He,et al.  Novel indirect enzyme-linked immunosorbent assay (ELISA) method to detect Total E. coli in water environment. , 2007, Analytica chimica acta.

[32]  R. Yazdanparast,et al.  Development and evaluation of a novel nucleic acid sequence-based amplification method using one specific primer and one degenerate primer for simultaneous detection of Salmonella Enteritidis and Salmonella Typhimurium. , 2013, Analytica chimica acta.