A rapid, sensitive and selective electrochemical biosensor with concanavalin A for the preemptive detection of norovirus.

Norovirus (NoV) is a foodborne pathogen that can cause sporadic and epidemic gastrointestinal diseases. Rapid screening is crucial to promptly identify the presence of NoV and prevent food poisoning. Here, we present a sensitive, selective, and rapid electrochemical biosensor for the detection of NoV. The proposed electrochemical biosensor is composed of a nanostructured gold electrode conjugated with concanavalin A (ConA). ConA functions as a recognition element that selectively captures NoV. Cyclic voltammetry revealed a linear relationship (R(2) = 0.998) between the current and concentration of NoV (in the range of 10(2) and 10(6) copies/mL), with a relatively short assay time (1h) and a good detection limit (35 copies/mL). Additionally, the signals of Hepatitis A and E in the selectively test were found to be only 2.0% and 2.8% of the NoV signal at an identical concentration of 10(3) copies/mL, proving that the electrochemical biosensor has a selectively of approximately 98%. Moreover, the concentration of NoV was measured in a realistic environment, i.e., a sample solution extracted from lettuce, to demonstrate a potential application of the proposed biosensor (LoD = 60 copies/mL).

[1]  M. Estes,et al.  Detection of Norwalk virus and hepatitis A virus in shellfish tissues with the PCR , 1995, Applied and environmental microbiology.

[2]  R. O'Kennedy,et al.  Advances in biosensors for detection of pathogens in food and water , 2003 .

[3]  P Tian,et al.  Detection of norovirus capsid proteins in faecal and food samples by a real time immuno‐PCR method , 2006, Journal of applied microbiology.

[4]  F. Smulders,et al.  Real-time PCR method with statistical analysis to compare the potential of DNA isolation methods to remove PCR inhibitors from samples for diagnostic PCR. , 2007, Molecular and cellular probes.

[5]  Duwoon Kim,et al.  Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus , 2014, Viruses.

[6]  D. D'Souza,et al.  Zirconium hydroxide effectively immobilizes and concentrates human enteric viruses , 2002, Letters in applied microbiology.

[7]  G. Palleschi,et al.  Comparison of PCR, Electrochemical Enzyme-Linked Immunosorbent Assays, and the Standard Culture Method for Detecting Salmonella in Meat Products , 2004, Applied and Environmental Microbiology.

[8]  P. D Patel,et al.  (Bio)sensors for measurement of analytes implicated in food safety: a review , 2002 .

[9]  R. Glass,et al.  Gastroenteritis viruses: an overview. , 2008, Novartis Foundation symposium.

[10]  Dmitri Ivnitski,et al.  Biosensors for detection of pathogenic bacteria , 1999 .

[11]  N. Sharon,et al.  Lectins as molecules and as tools. , 1986, Annual review of biochemistry.

[12]  Utkan Demirci,et al.  Quantum dot-based HIV capture and imaging in a microfluidic channel. , 2009, Biosensors & bioelectronics.

[13]  D. Heisey-Grove,et al.  An outbreak of norovirus gastroenteritis associated with wedding cakes , 2005, Epidemiology and Infection.

[14]  N. Ajami,et al.  Noroviruses: The leading cause of gastroenteritis worldwide. , 2010, Discovery medicine.

[15]  G. T. Pereira,et al.  Detection of infectious bronchitis virus and specific anti- viral antibodies using a Concanavalin A-Sandwich-ELISA. , 2005, Viral immunology.

[16]  M. Estes,et al.  Detection and analysis of a small round-structured virus strain in oysters implicated in an outbreak of acute gastroenteritis , 1996, Applied and environmental microbiology.

[17]  T. Ando,et al.  Western blot (immunoblot) assay of small, round-structured virus associated with an acute gastroenteritis outbreak in Tokyo , 1989, Journal of clinical microbiology.

[18]  James E. Robinson,et al.  Neutralizing and non-neutralizing monoclonal antibodies against dengue virus E protein derived from a naturally infected patient , 2010, Virology Journal.

[19]  Wenwen Tu,et al.  Electrochemical monitoring of an important biomarker and target protein: VEGFR2 in cell lysates , 2014, Scientific Reports.

[20]  G. Richards,et al.  Rapid and Efficient Extraction Method for Reverse Transcription-PCR Detection of Hepatitis A and Norwalk-Like Viruses in Shellfish , 2001, Applied and Environmental Microbiology.

[21]  Ilaria Palchetti,et al.  Electroanalytical biosensors and their potential for food pathogen and toxin detection , 2008, Analytical and bioanalytical chemistry.

[22]  J. Vinjé,et al.  International Collaborative Study To Compare Reverse Transcriptase PCR Assays for Detection and Genotyping of Noroviruses , 2003, Journal of Clinical Microbiology.

[23]  Yingchun Fu,et al.  Exploiting enzyme catalysis in ultra-low ion strength media for impedance biosensing of avian influenza virus using a bare interdigitated electrode. , 2014, Analytical chemistry.

[24]  M. Sobsey,et al.  Improved methods for detecting enteric viruses in oysters , 1978, Applied and environmental microbiology.

[25]  Richard G. Compton,et al.  The Use of Screen-Printed Electrodes in a Proof of Concept Electrochemical Estimation of Homocysteine and Glutathione in the Presence of Cysteine Using Catechol , 2014, Sensors.

[26]  Lele Zhang,et al.  Electrochemical measurement of Clostridium tetani using a reduced graphene oxide modified electrode and polyaniline–gold nanoparticle-labelled probe , 2014 .

[27]  M. Hur,et al.  Evaluation of Rapid Antigen Test for the Detection of Norovirus Infection: Comparison with ELISA and Real Time Quantitative Reverse Transcription PCR Assays. , 2011 .

[28]  S. Fukushi,et al.  Broadly Reactive and Highly Sensitive Assay for Norwalk-Like Viruses Based on Real-Time Quantitative Reverse Transcription-PCR , 2003, Journal of Clinical Microbiology.

[29]  Amethist S. Finch,et al.  A chemically synthesized capture agent enables the selective, sensitive, and robust electrochemical detection of anthrax protective antigen. , 2013, ACS nano.

[30]  Arunas Ramanavicius,et al.  Comparative study of surface plasmon resonance, electrochemical and electroassisted chemiluminescence methods based immunosensor for the determination of antibodies against human growth hormone. , 2012, Biosensors & bioelectronics.

[31]  D. Sano,et al.  Virus-Binding Proteins Recovered from Bacterial Culture Derived from Activated Sludge by Affinity Chromatography Assay Using a Viral Capsid Peptide , 2004, Applied and Environmental Microbiology.

[32]  M. Estes,et al.  Development of Methods To Detect “Norwalk-Like Viruses” (NLVs) and Hepatitis A Virus in Delicatessen Foods: Application to a Food-Borne NLV Outbreak , 2000, Applied and Environmental Microbiology.

[33]  D. Graham,et al.  Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein , 1992, Journal of virology.

[34]  T. Yokoyama,et al.  Varicella-zoster virus gH:gL contains a structure reactive with the anti-human gamma chain of IgG near the glycosylation site. , 2001, The Journal of general virology.

[35]  J. Brassard,et al.  An outbreak of norovirus caused by consumption of oysters from geographically dispersed harvest sites, British Columbia, Canada, 2004. , 2007, Foodborne pathogens and disease.

[36]  K. Evans-Nguyen,et al.  Protein arrays on patterned porous gold substrates interrogated with mass spectrometry: detection of peptides in plasma. , 2008, Analytical chemistry.

[37]  C. Fenselau,et al.  Lectin-based affinity capture for MALDI-MS analysis of bacteria. , 1999, Analytical chemistry.

[38]  Sung Yang,et al.  Rapid detection of norovirus from fresh lettuce using immunomagnetic separation and a quantum dots assay. , 2013, Journal of food protection.

[39]  Jan Vinjé,et al.  Noroviruses: a comprehensive review. , 2009, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[40]  C. Gaulin,et al.  Épidémie de gastro-entérite d'origine virale associée à la consommation de framboises importées , 1999 .

[41]  I. Seymour,et al.  Foodborne viruses and fresh produce , 2001, Journal of applied microbiology.

[42]  C. West,et al.  Identification of concanavalin A receptors and galactose-binding proteins in purified plasma membranes of Dictyostelium discoideum , 1977, The Journal of cell biology.

[43]  Maria C. DeRosa,et al.  Ultrasensitive Norovirus Detection Using DNA Aptasensor Technology , 2013, PloS one.

[44]  A. J. Bhattacharyya,et al.  Titania nanotube-modified screen printed carbon electrodes enhance the sensitivity in the electrochemical detection of proteins. , 2014, Bioelectrochemistry.

[45]  H. Bittiger,et al.  Concanavalin A as a tool , 1976 .

[46]  K. Kniel,et al.  Comparative recovery of foodborne viruses from fresh produce. , 2008, Foodborne pathogens and disease.

[47]  Harsh Sharma,et al.  Review of biosensors for foodborne pathogens and toxins , 2013 .

[48]  R. Glass,et al.  A foodborne outbreak of gastroenteritis associated with Norwalk-like viruses: first molecular traceback to deli sandwiches contaminated during preparation. , 2000, The Journal of infectious diseases.

[49]  M. Oh,et al.  Detection of hepatitis a virus from oyster by nested PCR using efficient extraction and concentration method , 2008, The Journal of Microbiology.

[50]  Yajiang Yin,et al.  Comparison between electrochemical ELISA and spectrophotometric ELISA for the detection of dentine sialophosphoprotein for root resorption. , 2014, American journal of orthodontics and dentofacial orthopedics : official publication of the American Association of Orthodontists, its constituent societies, and the American Board of Orthodontics.