A nanoparticle-based bio-barcode assay for ultrasensitive detection of ricin toxin.

The ultrasensitive bio-barcode amplification assay (BCA) technique was developed for the specific detection of the A chain of ricin toxin. The target antigen A chain was first captured by gold nanoparticles (GNPs) coated with polyclonal antibodies. Magnetic microparticles (MMPs) coated with A chain monoclonal antibody were then added to form a sandwich immuno-complex. After the immuno-complex was formed, signal DNA annealed to DNA strands covalently bound to the GNPs were released by heating and characterized by PCR and real-time fluorescence PCR. A detection limit of 1fg/ml was measured for A chain, six orders of magnitude more sensitive than that of conventional antigen-capture ELISA. The coefficient of variation (CV) of intra-assay and inter-assay ranged from 3.39% to 6.84%. The BCA can detect the A chain in milk and water mimic samples. In the following work it is demonstrated that this assay is a highly sensitive method for the detection of ricin proteins that could be adapted to measure other proteins.

[1]  John T McDevitt,et al.  Aptamer-based sensor arrays for the detection and quantitation of proteins. , 2004, Analytical chemistry.

[2]  W. Tolleson,et al.  A functional quantitative polymerase chain reaction assay for ricin, Shiga toxin, and related ribosome-inactivating proteins. , 2010, Analytical biochemistry.

[3]  L. Baert,et al.  Laboratory efforts to eliminate contamination problems in the real-time RT-PCR detection of noroviruses. , 2009, Journal of microbiological methods.

[4]  T. W. O'brien,et al.  Detection of ricin in food using electrochemiluminescence-based technology. , 2008, Journal of AOAC International.

[5]  C. Fournier-Wirth,et al.  Nanotechnologies for pathogen detection: Future alternatives? , 2010, Biologicals : journal of the International Association of Biological Standardization.

[6]  M. Poli,et al.  Detection of ricin by colorimetric and chemiluminescence ELISA. , 1994, Toxicon : official journal of the International Society on Toxinology.

[7]  Ronald M Atlas,et al.  Bioterriorism: from threat to reality. , 2002, Annual review of microbiology.

[8]  Silica coating magnetic nanoparticle-based silver enhancement immunoassay for rapid electrical detection of ricin toxin. , 2010, Toxicon : official journal of the International Society on Toxinology.

[9]  C. Mirkin,et al.  Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins , 2003, Science.

[10]  V. K. Rao,et al.  Amperometric immunosensor for ricin by using on graphite and carbon nanotube paste electrodes. , 2010, Talanta.

[11]  Subinoy Rana,et al.  Nanoparticles for detection and diagnosis. , 2010, Advanced drug delivery reviews.

[12]  C. Dogovski,et al.  Surface plasmon resonance detection of ricin and horticultural ricin variants in environmental samples. , 2008, Toxicon : official journal of the International Society on Toxinology.

[13]  C. Mirkin,et al.  Nanoparticle-based detection in cerebral spinal fluid of a soluble pathogenic biomarker for Alzheimer's disease. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Rory Stine,et al.  Comparison of glycosphingolipids and antibodies as receptor molecules for ricin detection. , 2005, Analytical chemistry.

[15]  Longyan Chen,et al.  Gold nanoparticle enhanced immuno-PCR for ultrasensitive detection of Hantaan virus nucleocapsid protein. , 2009, Journal of immunological methods.

[16]  Chad A Mirkin,et al.  Bio-bar-code-based DNA detection with PCR-like sensitivity. , 2004, Journal of the American Chemical Society.

[17]  Chad A Mirkin,et al.  The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange , 2006, Nature Protocols.

[18]  Yasuo Seto,et al.  A novel sugar-probe biosensor for the deadly plant proteinous toxin, ricin. , 2008, Biosensors & bioelectronics.

[19]  P. Thullier,et al.  Comparison of an electrochemiluminescence assay in plate format over a colorimetric ELISA, for the detection of ricin B chain (RCA-B). , 2007, Journal of immunological methods.

[20]  Daniel C. Smith,et al.  Ricin: the endoplasmic reticulum connection. , 2004, Toxicon : official journal of the International Society on Toxinology.

[21]  Aptamer-based bio-barcode assay for the detection of cytochrome-c released from apoptotic cells. , 2010, Biochemical and biophysical research communications.

[22]  E. Alocilja,et al.  Fluorescent bio-barcode DNA assay for the detection of Salmonella enterica serovar Enteritidis. , 2009, Biosensors & bioelectronics.

[23]  U. Narang,et al.  Fiber optic-based biosensor for ricin. , 1997, Biosensors & bioelectronics.

[24]  Chad A Mirkin,et al.  A fluorophore-based bio-barcode amplification assay for proteins. , 2006, Small.

[25]  A Pihl,et al.  Radioimmunoassays of abrin and ricin in blood. , 1981, Journal of toxicology and environmental health.