Monitoring microbial populations of sulfate-reducing bacteria using an impedimetric immunosensor based on agglutination assay.

An impedimetric immunosensor was fabricated for rapid and non-labeled detection of sulfate-reducing bacteria, Desulforibrio caledoiensis (SRB) by immobilizing lectin-Concanavalin A using an agglutination assay. The immobilization of lectin was conducted using amine coupling on the surface of a gold (Au) electrode assembled with 11-Mercaptoundecanoic acid. Electrochemical impedance spectroscopy (EIS) was used to verify the stepwise assembly of the sensor system. The work conditions of the impedimetric immunosensor, such as pH of the buffer solutions and the incubation time of lectin, were optimized. Faradic impedance spectra for charge transfer for the redox probe Fe(CN)(6)(3-/4-)were measured to determine SRB concentrations. The diameter of the Nyquist diagram that is equal to the charge-transfer resistance (R(ct)) increased with increasing SRB concentration. A linear relationship between R(ct) and SRB concentration was obtained in SRB concentration range of 1.8 to 1.8 x 10(7)cfu/ml. The variation of the SRB population during the growth process was also monitored using the impedimetric immunosensor. This approach has great potential for simple, low-cost, and time-saving monitoring of microbial populations.

[1]  Yongcheng Liu,et al.  Detection of pathogens using luminescent CdSe/ZnS dendron nanocrystals and a porous membrane immunofilter. , 2007, Analytical chemistry.

[2]  Michael Keusgen,et al.  Magnetic biosensor for the detection of Yersinia pestis. , 2007, Journal of microbiological methods.

[3]  Hsien-Chang Chang,et al.  Impedance spectral studies of self-assembly of alkanethiols with different chain lengths using different immobilization strategies on Au electrodes , 2005 .

[4]  Y. Abd-el-Malek,et al.  Counting of Sulphate-reducing Bacteria in Mixed Bacterial Populations , 1958, Nature.

[5]  Xiangqun Zeng,et al.  Nonlabeled quartz crystal microbalance biosensor for bacterial detection using carbohydrate and lectin recognitions. , 2007, Analytical chemistry.

[6]  Yu-Chie Chen,et al.  Using biofunctionalized nanoparticles to probe pathogenic bacteria. , 2004, Analytical chemistry.

[7]  Yu-Chie Chen,et al.  Affinity capture using vancomycin-bound magnetic nanoparticles for the MALDI-MS analysis of bacteria. , 2005, Analytical chemistry.

[8]  Chih-Ching Huang,et al.  Synthesis of fluorescent carbohydrate-protected Au nanodots for detection of Concanavalin A and Escherichia coli. , 2009, Analytical chemistry.

[9]  N. Sharon,et al.  Lectins: Carbohydrate-Specific Proteins That Mediate Cellular Recognition. , 1998, Chemical reviews.

[10]  Y. Fung,et al.  Self-assembled monolayers as the coating in a quartz piezoelectric crystal immunosensor to detect Salmonella in aqueous solution. , 2001, Analytical chemistry.

[11]  Michael Wagner,et al.  Improved 16S rRNA-targeted probe set for analysis of sulfate-reducing bacteria by fluorescence in situ hybridization. , 2007, Journal of microbiological methods.

[12]  T. Montag,et al.  Bacteria detection by flow cytometry , 2008, Clinical chemistry and laboratory medicine.

[13]  Bengt Danielsson,et al.  Flow-injection assay of the pathogenic bacteria using lectin-based quartz crystal microbalance biosensor , 2008 .

[14]  C. Gaylarde,et al.  New rapid methods for the identification of sulphate-reducing bacteria. , 1990 .

[15]  Y. Fung,et al.  Nano-silver-modified PQC/DNA biosensor for detecting E. coli in environmental water. , 2009, Biosensors & bioelectronics.

[16]  Da-Jeng Yao,et al.  Magnetic bead-based DNA detection with multi-layers quantum dots labeling for rapid detection of Escherichia coli O157:H7. , 2008, Biosensors & bioelectronics.

[17]  Y. Chuang,et al.  Disposable amperometric immunosensing strips fabricated by Au nanoparticles-modified screen-printed carbon electrodes for the detection of foodborne pathogen Escherichia coli O157:H7. , 2008, Biosensors & bioelectronics.

[18]  Hsieh-Cheng Han,et al.  Application of parylene-coated quartz crystal microbalance for on-line real-time detection of microbial populations. , 2009, Biosensors & bioelectronics.

[19]  Yanbin Li,et al.  Immunobiosensor chips for detection of Escherichia coil O157:H7 using electrochemical impedance spectroscopy. , 2002, Analytical chemistry.

[20]  Michael Keusgen,et al.  Rapid method for detection of Salmonella in milk by surface plasmon resonance (SPR). , 2007, Biosensors & bioelectronics.

[21]  Xiuheng Xue,et al.  Fluorescence detection of total count of Escherichia coli and Staphylococcus aureus on water-soluble CdSe quantum dots coupled with bacteria. , 2009, Talanta.

[22]  H. Angerstein-Kozlowska,et al.  Elementary steps of electrochemical oxidation of single-crystal planes of Au—I. Chemical basis of processes involving geometry of anions and the electrode surfaces , 1986 .

[23]  Li Yan,et al.  Effects of sulfate-reducing bacteria on the corrosion behavior of carbon steel , 2007 .

[24]  I. Suni,et al.  Impedance biosensor for peanut protein Ara h 1. , 2008, Analytical chemistry.

[25]  Marek Piliarik,et al.  High-throughput SPR sensor for food safety. , 2009, Biosensors & bioelectronics.

[26]  Yuxiao Cheng,et al.  Combining biofunctional magnetic nanoparticles and ATP bioluminescence for rapid detection of Escherichia coli. , 2009, Talanta.

[27]  A. Gehring,et al.  Immunoelectrochemical assays for bacteria: use of epifluorescence microscopy and rapid-scan electrochemical techniques in development of an assay for Salmonella. , 1996, Analytical chemistry.

[28]  M. Cotta,et al.  Evaluation of the sulfate-reducing bacterial population associated with stored swine slurry. , 2008, Anaerobe.

[29]  Zirong Wu,et al.  Self-assembled monolayers-based immunosensor for detection of Escherichia coli using electrochemical impedance spectroscopy , 2008 .

[30]  Vincent M Rotello,et al.  Rapid and efficient identification of bacteria using gold-nanoparticle-poly(para-phenyleneethynylene) constructs. , 2008, Angewandte Chemie.

[31]  Tit Meng Lim,et al.  Detection of Saccharomyces cerevisiae immobilized on self-assembled monolayer (SAM) of alkanethiolate using electrochemical impedance spectroscopy , 2005 .

[32]  K. Ingvorsen,et al.  Improved Most-Probable-Number Method To Detect Sulfate-Reducing Bacteria with Natural Media and a Radiotracer , 1998, Applied and Environmental Microbiology.