An improved sensitive assay for the detection of PSP toxins with neuroblastoma cell-based impedance biosensor.

Paralytic shellfish poisoning (PSP) toxins are well-known sodium channel-blocking marine toxins, which block the conduction of nerve impulses and lead to a series of neurological disorders symptoms. However, PSP toxins can inhibit the cytotoxicity effect of compounds (e.g., ouabain and veratridine). Under the treatment of ouabain and veratridine, neuroblastoma cell will swell and die gradually, since veratridine causes the persistent inflow of Na(+) and ouabain inhibits the activity of Na(+)/K(+)-ATPases. Therefore, PSP toxins with antagonism effect can raise the chance of cell survival by blocking inflow of Na(+). Based on the antagonism effect of PSP toxins, we designed an improved cell-based assay to detect PSP toxins using a neuroblastoma cell-based impedance biosensor. The results demonstrated that this biosensor showed high sensitivity and good specificity for saxitoxins detection. The detection limit of this biosensor was as low as 0.03 ng/ml, which was lower than previous reported cell-based assays and mouse bioassays. With the improvement of biosensor performance, the neuroblastoma cell-based impedance biosensor has great potential to be a universal PSP screening method.

[1]  N. Hu,et al.  Assessment of cadmium-induced hepatotoxicity and protective effects of zinc against it using an improved cell-based biosensor , 2013 .

[2]  Krista Thomas,et al.  Comparison of AOAC 2005.06 LC official method with other methodologies for the quantitation of paralytic shellfish poisoning toxins in UK shellfish species , 2011, Analytical and bioanalytical chemistry.

[3]  E. Moore,et al.  Comparison of Cell-Based Biosensors with Traditional Analytical Techniques for Cytotoxicity Monitoring and Screening of Polycyclic Aromatic Hydrocarbons in the Environment , 2009 .

[4]  A. Verma,et al.  Ras mutation, irrespective of cell type and p53 status, determines a cell's destiny to undergo apoptosis by okadaic acid, an inhibitor of protein phosphatase 1 and 2A. , 1999, Molecular pharmacology.

[5]  I. Rietjens,et al.  Marine neurotoxins: state of the art, bottlenecks, and perspectives for mode of action based methods of detection in seafood. , 2014, Molecular nutrition & food research.

[6]  M. Kane,et al.  A cytotoxicity assay for the detection and differentiation of two families of shellfish toxins. , 2001, Toxicon : official journal of the International Society on Toxinology.

[7]  Katrina Campbell,et al.  Development and validation of an ultrasensitive fluorescence planar waveguide biosensor for the detection of paralytic shellfish toxins in marine algae. , 2013, Biosensors & bioelectronics.

[8]  Luis M. Botana,et al.  Seafood and freshwater toxins : pharmacology, physiology, and detection , 2000 .

[9]  Jean-Louis Marty,et al.  Biosensors to detect marine toxins: Assessing seafood safety. , 2007, Talanta.

[10]  Z. Kang,et al.  Tracing the origin of paralytic shellfish toxins in scallop Patinopecten yessoensis in the northern Yellow Sea , 2013, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[11]  Kaiqi Su,et al.  A cardiomyocyte-based biosensor for antiarrhythmic drug evaluation by simultaneously monitoring cell growth and beating. , 2013, Biosensors & bioelectronics.

[12]  A. Malik,et al.  Electrical method for detection of endothelial cell shape change in real time: assessment of endothelial barrier function. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[13]  K Kogure,et al.  A tissue culture assay for tetrodotoxin, saxitoxin and related toxins. , 1988, Toxicon : official journal of the International Society on Toxinology.

[14]  Liju Yang,et al.  Real-time electrical impedance detection of cellular activities of oral cancer cells. , 2010, Biosensors & bioelectronics.

[15]  M. de Carvalho,et al.  Paralytic shellfish poisoning due to ingestion of Gymnodinium catenatum contaminated cockles--application of the AOAC HPLC official method. , 2012, Toxicon : official journal of the International Society on Toxinology.

[16]  Fumio Kondo,et al.  Cell bioassay for paralytic shellfish poisoning (PSP): comparison with postcolumn derivatization liquid chromatographic analysis and application to the monitoring of PSP in shellfish. , 2006, Journal of agricultural and food chemistry.

[17]  Stacey M Etheridge,et al.  Paralytic shellfish poisoning: seafood safety and human health perspectives. , 2010, Toxicon : official journal of the International Society on Toxinology.

[18]  M. Wekell,et al.  Detection of paralytic shellfish poison by rapid cell bioassay: antagonism of voltage-gated sodium channel active toxins in vitro. , 2003, Journal of AOAC International.

[19]  Gary S. Sayler,et al.  An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions , 2013, Marine drugs.

[20]  A. Gago-Martínez,et al.  Extension of the validation of AOAC Official Method 2005.06 for dc-GTX2,3: interlaboratory study. , 2012, Journal of AOAC International.

[21]  H. Tsuzuki,et al.  A rapid detection method for paralytic shellfish poisoning toxins by cell bioassay. , 2005, Toxicon : official journal of the International Society on Toxinology.

[22]  J. Diogène,et al.  Comparative study of the use of neuroblastoma cells (Neuro-2a) and neuroblastomaxglioma hybrid cells (NG108-15) for the toxic effect quantification of marine toxins. , 2008, Toxicon : official journal of the International Society on Toxinology.

[23]  R. Nicholson,et al.  A rapid and sensitive assay for paralytic shellfish poison (PSP) toxins using mouse brain synaptoneurosomes. , 2002, Toxicon : official journal of the International Society on Toxinology.

[24]  A. Humpage,et al.  Comparison of analytical tools and biological assays for detection of paralytic shellfish poisoning toxins , 2010, Analytical and bioanalytical chemistry.

[25]  L. Botana,et al.  A fluorimetric microplate assay for detection and quantitation of toxins causing paralytic shellfish poisoning. , 2003, Chemical research in toxicology.

[26]  N. Hu,et al.  Comparison between ECIS and LAPS for establishing a cardiomyocyte-based biosensor , 2013 .

[27]  Qingjun Liu,et al.  Impedance studies of bio-behavior and chemosensitivity of cancer cells by micro-electrode arrays. , 2009, Biosensors & bioelectronics.

[28]  Qingjun Liu,et al.  A novel microphysiometer based on high sensitivity LAPS and microfluidic system for cellular metabolism study and rapid drug screening. , 2013, Biosensors & bioelectronics.

[29]  M. Wiese,et al.  Neurotoxic Alkaloids: Saxitoxin and Its Analogs , 2010, Marine drugs.

[30]  N. Kulagina,et al.  Pharmacological effects of the marine toxins, brevetoxin and saxitoxin, on murine frontal cortex neuronal networks. , 2004, Toxicon : official journal of the International Society on Toxinology.

[31]  I. Cree Cancer Cell Culture , 2011, Methods in Molecular Biology.

[32]  A. Takai,et al.  Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics. , 1988, The Biochemical journal.

[33]  Zheng Xiaoxiang,et al.  Drug evaluations using a novel microphysiometer based on cell-based biosensors , 2001 .

[34]  Miqin Zhang,et al.  Cellular impedance biosensors for drug screening and toxin detection. , 2007, The Analyst.