Centrifugal microfluidic platform for ultrasensitive detection of botulinum toxin

Koh, Chung -Yan; Schaff, Ulrich Y.; Piccini, Matthew E.; Stanker, Larry H.; Cheng, Luisa W.; Ravichandran, Easwaran; Singh, Bal -Ram; Sommer, Greg J.; Singh, Anup K.

doi:10.1021/ac504054u

Centrifugal microfluidic platform for ultrasensitive detection of botulinum toxin

Journal Article · Thu Dec 18 00:00:00 EST 2014 · Analytical Chemistry

DOI:https://doi.org/10.1021/ac504054u· OSTI ID:1184997

Koh, Chung -Yan ^[1]; Schaff, Ulrich Y. ^[2]; Piccini, Matthew E. ^[3]; Stanker, Larry H. ^[4]; Cheng, Luisa W. ^[4]; Ravichandran, Easwaran ^[5]; Singh, Bal -Ram ^[5]; Sommer, Greg J. ^[2]; Singh, Anup K. ^[1]

Sandia National Lab. (SNL-CA), Livermore, CA (United States)
Sandia National Lab. (SNL-CA), Livermore, CA (United States); Sandstone Diagnostics, Livermore, CA (United States)
Sandia National Lab. (SNL-CA), Livermore, CA (United States); Cepheid, Sunnyvale, CA (United States)
U.S. Dept. of Agriculture - Agricultural Research Service, Albany, CA (United States)
Univ. of Massachusetts, Dartmouth, MA (United States)

In this study, we present an innovative centrifugal microfluidic immunoassay platform (SpinDx) to address the urgent biodefense and public health need for ultrasensitive point-of-care/incident detection of botulinum toxin. The simple, sample-to-answer centrifugal microfluidic immunoassay approach is based on binding of toxins to antibody-laden capture particles followed by sedimentation of the particles through a density-media in a microfluidic disk and quantification by laser-induced fluorescence. A blind, head-to-head comparison study of SpinDx versus the gold-standard mouse bioassay demonstrates 100-fold improvement in sensitivity (limit of detection = 0.09 pg/mL), while achieving total sample-to-answer time of <30 min with 2-μL required volume of the unprocessed sample. We further demonstrate quantification of botulinum toxin in both exogeneous (human blood and serum spiked with toxins) and endogeneous (serum from mice intoxicated via oral, intranasal, and intravenous routes) samples. SpinDx can analyze, without any sample preparation, multiple sample types including whole blood, serum, and food. It is readily expandable to additional analytes as the assay reagents (i.e., the capture beads and detection antibodies) are disconnected from the disk architecture and the reader, facilitating rapid development of new assays. SpinDx can also serve as a general-purpose immunoassay platform applicable to diagnosis of other conditions and diseases.

View Accepted Manuscript (DOE)

Research Organization:: Sandia National Laboratories (SNL-CA), Livermore, CA (United States)

Sponsoring Organization:: NIH

Grant/Contract Number:: AC04-94AL85000

OSTI ID:: 1184997

Report Number(s):: SAND--2014-20313J; 547681

Journal Information:: Analytical Chemistry, Journal Name: Analytical Chemistry Journal Issue: 2 Vol. 87; ISSN 0003-2700

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

References (53)

Mouse diaphragm assay for detection of antibodies against botulinum toxin type B Dressler, Dirk; Lange, M.; Bigalke, Hans Movement Disorders, Vol. 20, Issue 12 https://doi.org/10.1002/mds.20625	journal	August 2005
Llama-derived single-domain antibodies for the detection of botulinum A neurotoxin Swain, Marla D.; Anderson, George P.; Zabetakis, Dan Analytical and Bioanalytical Chemistry, Vol. 398, Issue 1 https://doi.org/10.1007/s00216-010-3905-3	journal	June 2010
Rapid detection of Clostridium botulinum toxins A, B, E, and F in clinical samples, selected food matrices, and buffer using paramagnetic bead-based electrochemiluminescence detection Rivera, Victor R.; Gamez, Frank J.; Keener, William K. Analytical Biochemistry, Vol. 353, Issue 2 https://doi.org/10.1016/j.ab.2006.02.030	journal	June 2006
Detection of six serotypes of botulinum neurotoxin using fluorogenic reporters Ruge, Daniel R.; Dunning, F. Mark; Piazza, Timothy M. Analytical Biochemistry, Vol. 411, Issue 2 https://doi.org/10.1016/j.ab.2011.01.002	journal	April 2011
Biosensing enhancement of dengue virus using microballoon mixers on centrifugal microfluidic platforms Aeinehvand, Mohammad Mahdi; Ibrahim, Fatimah; Harun, Sulaiman Wadi Biosensors and Bioelectronics, Vol. 67 https://doi.org/10.1016/j.bios.2014.08.076	journal	May 2015
Towards an international standard for detection and typing botulinum neurotoxin-producing Clostridia types A, B, E and F in food, feed and environmental samples: A European ring trial study to evaluate a real-time PCR assay Fenicia, Lucia; Fach, Patrick; van Rotterdam, Bart J. International Journal of Food Microbiology, Vol. 145 https://doi.org/10.1016/j.ijfoodmicro.2011.02.001	journal	March 2011
Development and partial characterization of high-affinity monoclonal antibodies for botulinum toxin type A and their use in analysis of milk by sandwich ELISA Stanker, Larry H.; Merrill, Paul; Scotcher, Miles C. Journal of Immunological Methods, Vol. 336, Issue 1 https://doi.org/10.1016/j.jim.2008.03.003	journal	July 2008
Medical aspects of bio-terrorism Balali-Mood, Mahdi; Moshiri, Mohammad; Etemad, Leila Toxicon, Vol. 69 https://doi.org/10.1016/j.toxicon.2013.01.005	journal	July 2013
Design of a Compact Disk-like Microfluidic Platform for Enzyme-Linked Immunosorbent Assay Lai, Siyi; Wang, Shengnian; Luo, Jun Analytical Chemistry, Vol. 76, Issue 7 https://doi.org/10.1021/ac0348322	journal	April 2004
Thermostable Llama Single Domain Antibodies for Detection of Botulinum A Neurotoxin Complex Goldman, Ellen R.; Anderson, George P.; Conway, Jerry Analytical Chemistry, Vol. 80, Issue 22 https://doi.org/10.1021/ac8014774	journal	November 2008
Simultaneous Multiplex Detection and Confirmation of the Proteinaceous Toxins Abrin, Ricin, Botulinum Toxins, and Staphylococcus Enterotoxins A, B, and C in Food Garber, Eric A. E.; Venkateswaran, Kodumudi V.; O’Brien, Thomas W. Journal of Agricultural and Food Chemistry, Vol. 58, Issue 11 https://doi.org/10.1021/jf100789n	journal	May 2010
A liposome-PCR assay for the ultrasensitive detection of biological toxins Mason, Jeffrey T.; Xu, Lixin; Sheng, Zong-mei Nature Biotechnology, Vol. 24, Issue 5 https://doi.org/10.1038/nbt1201	journal	April 2006
A fully automated immunoassay from whole blood on a disc Lee, Beom Seok; Lee, Jung-Nam; Park, Jong-Myeon Lab on a Chip, Vol. 9, Issue 11, p. 1548-1555 https://doi.org/10.1039/b820321k	journal	January 2009
Centrifugal microfluidics for biomedical applications Gorkin, Robert; Park, Jiwoon; Siegrist, Jonathan Lab on a Chip, Vol. 10, Issue 14, p. 1758-1773 https://doi.org/10.1039/b924109d	journal	January 2010
A self-pumping lab-on-a-chip for rapid detection of botulinum toxin Lillehoj, Peter B.; Wei, Fang; Ho, Chih-Ming Lab on a Chip, Vol. 10, Issue 17, p. 2265-2270 https://doi.org/10.1039/c004885b	journal	July 2010
Adopting Orphan Drugs — Two Dozen Years of Treating Rare Diseases Haffner, Marlene E. New England Journal of Medicine, Vol. 354, Issue 5 https://doi.org/10.1056/nejmp058317	journal	February 2006
Botulism Diagnostics: From Clinical Symptoms to in vitro Assays Cai, Shuowei; Singh, Bal Ram; Sharma, Shashi Critical Reviews in Microbiology, Vol. 33, Issue 2 https://doi.org/10.1080/10408410701364562	journal	January 2007
Application of Real-Time PCR for Quantitative Detection of Clostridium botulinum Type A Toxin Gene in Food Yoon, So-Yeon; Chung, Gyung Tae; Kang, Do-Hyun Microbiology and Immunology, Vol. 49, Issue 6 https://doi.org/10.1111/j.1348-0421.2005.tb03755.x	journal	June 2005
Laboratory Diagnostics of Botulism Lindström, Miia; Korkeala, Hannu Clinical Microbiology Reviews, Vol. 19, Issue 2 https://doi.org/10.1128/cmr.19.2.298-314.2006	journal	April 2006
Analysis of the Mechanisms That Underlie Absorption of Botulinum Toxin by the Inhalation Route Al-Saleem, Fetweh H.; Ancharski, Denise M.; Joshi, Suresh G. Infection and Immunity, Vol. 80, Issue 12 https://doi.org/10.1128/iai.00669-12	journal	September 2012
Bacterial toxins: a table of lethal amounts Gill, D. M. Microbiological Reviews, Vol. 46, Issue 1 https://doi.org/10.1128/mr.46.1.86-94.1982	journal	March 1982
Properties and use of botulinum toxin and other microbial neurotoxins in medicine Schantz, E. J.; Johnson, E. A. Microbiological Reviews, Vol. 56, Issue 1 https://doi.org/10.1128/mr.56.1.80-99.1992	journal	March 1992
Lab On A CD Madou, Marc; Zoval, Jim; Jia, Guangyao Annual Review of Biomedical Engineering, Vol. 8, Issue 1, p. 601-628 https://doi.org/10.1146/annurev.bioeng.8.061505.095758	journal	August 2006
Bioterrorism in Canada: An Economic Assessment of Prevention and Postattack Response John, Ronald St; Finlay, Brian; Blair, Curtis Canadian Journal of Infectious Diseases, Vol. 12, Issue 5 https://doi.org/10.1155/2001/904148	journal	January 2001
Real-time polymerase chain reaction for the detection of toxigenic Clostridium botulinum type C1 in waterbird and sediment samples: comparison with other PCR techniques Vidal, Dolors; Taggart, Mark A.; Badiola, Ignacio Journal of Veterinary Diagnostic Investigation, Vol. 23, Issue 5 https://doi.org/10.1177/1040638711416847	journal	September 2011
Universal and specific quantitative detection of botulinum neurotoxin genes Hill, Brenna J.; Skerry, Janet C.; Smith, Theresa J. BMC Microbiology, Vol. 10, Issue 1 https://doi.org/10.1186/1471-2180-10-267	journal	January 2010
Attomolar Detection of Botulinum Toxin Type A in Complex Biological Matrices Bagramyan, Karine; Barash, Jason R.; Arnon, Stephen S. PLoS ONE, Vol. 3, Issue 4 https://doi.org/10.1371/journal.pone.0002041	journal	April 2008
Detection of Botulinum Neurotoxin Serotype B at Sub Mouse LD50 Levels by a Sandwich Immunoassay and Its Application to Toxin Detection in Milk Scotcher, Miles C.; Cheng, Luisa W.; Stanker, Larry H. PLoS ONE, Vol. 5, Issue 6 https://doi.org/10.1371/journal.pone.0011047	journal	June 2010
Whole Blood Immunoassay Based on Centrifugal Bead Sedimentation Schaff, U. Y.; Sommer, G. J. Clinical Chemistry, Vol. 57, Issue 5, p. 753-761 https://doi.org/10.1373/clinchem.2011.162206	journal	March 2011
The Gyrolab™ immunoassay system: a platform for automated bioanalysis and rapid sample turnaround Fraley, Kristy J.; Abberley, Lee; Hottenstein, Charles S. Bioanalysis, Vol. 5, Issue 14 https://doi.org/10.4155/bio.13.145	journal	July 2013
Methods for Detection of Clostridium botulinum Toxin in Foods† Sharma, Shashi K.; Whiting, Richard C. Journal of Food Protection, Vol. 68, Issue 6 https://doi.org/10.4315/0362-028x-68.6.1256	journal	June 2005
Botulinum Toxin as a Biological Weapon: Medical and Public Health Management Arnon, Stephen S.; Schechter, Robert; Inglesby, Thomas V. JAMA, Vol. 285, Issue 8 https://doi.org/10.1001/jama.285.8.1059	journal	February 2001
Ultrasensitive Detection of Botulinum Neurotoxins and Anthrax Lethal Factor in Biological Samples by ALISSA Bagramyan, Karine; Kalkum, Markus Microbial Toxins https://doi.org/10.1007/978-1-61779-102-4_3	book	January 2011
Improved detection of botulinum neurotoxin type A in stool by mass spectrometry Wang, Dongxia; Baudys, Jakub; Kalb, Suzanne R. Analytical Biochemistry, Vol. 412, Issue 1 https://doi.org/10.1016/j.ab.2011.01.025	journal	May 2011
Detection of botulinum neurotoxin-A activity in food by peptide cleavage assay Rasooly, R.; Stanker, L.; Carter, J. International Journal of Food Microbiology, Vol. 126, Issue 1-2 https://doi.org/10.1016/j.ijfoodmicro.2008.05.012	journal	August 2008
Rapid and selective detection of botulinum neurotoxin serotype-A and -B with a single immunochromatographic test strip Ching, Kathryn H.; Lin, Alice; McGarvey, Jeffery A. Journal of Immunological Methods, Vol. 380, Issue 1-2 https://doi.org/10.1016/j.jim.2012.03.008	journal	June 2012
Comparison of oral toxicological properties of botulinum neurotoxin serotypes A and B Cheng, Luisa W.; Henderson, Thomas D. Toxicon, Vol. 58, Issue 1 https://doi.org/10.1016/j.toxicon.2011.05.003	journal	July 2011
Gangliosides as Receptors for Biological Toxins: Development of Sensitive Fluoroimmunoassays Using Ganglioside-Bearing Liposomes Singh, Anup K.; Harrison, Suzanne H.; Schoeniger, Joseph S. Analytical Chemistry, Vol. 72, Issue 24 https://doi.org/10.1021/ac000846l	journal	December 2000
Antibody Quantum Dot Conjugates Developed via Copper-Free Click Chemistry for Rapid Analysis of Biological Samples Using a Microfluidic Microsphere Array System Kotagiri, Nalinikanth; Li, Zhenyu; Xu, Xiaoxiao Bioconjugate Chemistry, Vol. 25, Issue 7 https://doi.org/10.1021/bc500139u	journal	June 2014
Simple Conjugation and Purification of Quantum Dot−Antibody Complexes Using a Thermally Responsive Elastin-Protein L Scaffold As Immunofluorescent Agents Lao, U. Loi; Mulchandani, Ashok; Chen, Wilfred Journal of the American Chemical Society, Vol. 128, Issue 46 https://doi.org/10.1021/ja065343x	journal	November 2006
Highly Sensitive Single Domain Antibody–Quantum Dot Conjugates for Detection of HER2 Biomarker in Lung and Breast Cancer Cells Rakovich, Tatsiana Y.; Mahfoud, Omar K.; Mohamed, Bashir M. ACS Nano, Vol. 8, Issue 6 https://doi.org/10.1021/nn500212h	journal	May 2014
Rapid and highly sensitive luciferase reporter assay for the automated detection of botulinum toxin in the centrifugal microfluidic LabDisk platform van Oordt, Thomas; Stevens, Gregory B.; Vashist, Sandeep Kumar RSC Advances, Vol. 3, Issue 44 https://doi.org/10.1039/c3ra44482a	journal	January 2013
Human Botulism Immune Globulin for the Treatment of Infant Botulism Arnon, Stephen S.; Schechter, Robert; Maslanka, Susan E. New England Journal of Medicine, Vol. 354, Issue 5 https://doi.org/10.1056/NEJMoa051926	journal	February 2006
Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification of Ricin in Suspicious White Powders and Environmental Samples Hodge, David R.; Prentice, Kristin Willner; Ramage, Jason G. Biosecurity and Bioterrorism: Biodefense Strategy, Practice, and Science, Vol. 11, Issue 4 https://doi.org/10.1089/bsp.2013.0053	journal	December 2013
Detection and Confirmation of Clostridium botulinum in Water Used for Cooling at a Plant Producing Low-Acid Canned Foods Sachdeva, Amita; Defibaugh-Chávez, Stephanie L. H.; Day, James B. Applied and Environmental Microbiology, Vol. 76, Issue 22 https://doi.org/10.1128/AEM.00820-10	journal	November 2010
Laboratory Diagnostics of Botulism Lindström, Miia; Korkeala, Hannu Clinical Microbiology Reviews, Vol. 19, Issue 2 https://doi.org/10.1128/CMR.19.2.298-314.2006	journal	April 2006
Analysis of the Mechanisms That Underlie Absorption of Botulinum Toxin by the Inhalation Route Al-Saleem, Fetweh H.; Ancharski, Denise M.; Joshi, Suresh G. Infection and Immunity, Vol. 80, Issue 12 https://doi.org/10.1128/IAI.00669-12	journal	September 2012
Oral toxicities of Clostridium botulinum type A and B toxins from different strains Ohishi, I. Infection and Immunity, Vol. 43, Issue 2 https://doi.org/10.1128/IAI.43.2.487-490.1984	journal	February 1984
Bacterial toxins: a table of lethal amounts. Gill, D. M. Microbiological Reviews, Vol. 46, Issue 1 https://doi.org/10.1128/MMBR.46.1.86-94.1982	journal	January 1982
Properties and use of botulinum toxin and other microbial neurotoxins in medicine. Schantz, E. J.; Johnson, E. A. Microbiological Reviews, Vol. 56, Issue 1 https://doi.org/10.1128/MMBR.56.1.80-99.1992	journal	January 1992
Oral toxicities of Clostridium botulinum type A and B toxins from different strains Ohishi, I. Infection and Immunity, Vol. 43, Issue 2 https://doi.org/10.1128/iai.43.2.487-490.1984	journal	February 1984
Current strategies for designing antidotes against botulinum neurotoxins Patel, Kruti; Cai, Shuowei; Singh, Bal Ram Expert Opinion on Drug Discovery, Vol. 9, Issue 3 https://doi.org/10.1517/17460441.2014.884066	journal	February 2014
Methods for Detection of Clostridium botulinum Toxin in Foods† Sharma, Shashi K.; Whiting, Richard C. Journal of Food Protection, Vol. 68, Issue 6 https://doi.org/10.4315/0362-028X-68.6.1256	journal	June 2005

Cited By (16)

A Rapid, Sensitive, and Portable Biosensor Assay for the Detection of Botulinum Neurotoxin Serotype A in Complex Food Matrices Tam, Christina C.; Flannery, Andrew R.; Cheng, Luisa W. Preprints.org https://doi.org/10.20944/preprints201808.0390.v1	posted_content	August 2018
Rapid, Portable, Multiplexed Detection of Bacterial Pathogens Directly from Clinical Sample Matrices Phaneuf, Christopher; Mangadu, Betty; Piccini, Matthew Biosensors, Vol. 6, Issue 4 https://doi.org/10.3390/bios6040049	journal	September 2016
A Review of Biomedical Centrifugal Microfluidic Platforms Tang, Minghui; Wang, Guanghui; Kong, Siu-Kai Micromachines, Vol. 7, Issue 2 https://doi.org/10.3390/mi7020026	journal	February 2016
Fabrication of a Novel Highly Sensitive and Selective Immunosensor for Botulinum Neurotoxin Serotype A Based on an Effective Platform of Electrosynthesized Gold Nanodendrites/Chitosan Nanoparticles Sorouri, Rahim; Bagheri, Hasan; Afkhami, Abbas Sensors, Vol. 17, Issue 5 https://doi.org/10.3390/s17051074	journal	May 2017
Rapid Detection of Botulinum Neurotoxins—A Review Hobbs, Robert J.; Thomas, Carol A.; Halliwell, Jennifer Toxins, Vol. 11, Issue 7 https://doi.org/10.3390/toxins11070418	journal	July 2019
Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test Worbs, Sylvia; Fiebig, Uwe; Zeleny, Reinhard Toxins, Vol. 7, Issue 12 https://doi.org/10.3390/toxins7124857	journal	November 2015
Advances in Reagents Storage and Release in Self‐Contained Point‐of‐Care Devices Deng, Jinqi; Jiang, Xingyu Advanced Materials Technologies, Vol. 4, Issue 6 https://doi.org/10.1002/admt.201800625	journal	January 2019
Microfluidics‐Implemented Biochemical Assays: From the Perspective of Readout Chen, Wenwen; Shao, Fangchi; Xianyu, Yunlei Small, Vol. 16, Issue 9 https://doi.org/10.1002/smll.201903388	journal	September 2019
Microfluidics for Biomedical Analysis Yang, Yong; Chen, Yong; Tang, Hao Small Methods, Vol. 4, Issue 4 https://doi.org/10.1002/smtd.201900451	journal	April 2020
Ultrasensitive multi-species detection of CRISPR-Cas9 by a portable centrifugal microfluidic platform Phaneuf, Christopher R.; Seamon, Kyle J.; Eckles, Tyler P. Analytical Methods, Vol. 11, Issue 5 https://doi.org/10.1039/c8ay02726a	journal	January 2019
A novel electromagnet-triggered pillar valve and its application in immunoassay on a centrifugal platform Chen, Yiqi; Shen, Minjie; Zhu, Yunzeng Lab on a Chip, Vol. 19, Issue 10 https://doi.org/10.1039/c9lc00043g	journal	January 2019
Applications of antibodies in microfluidics-based analytical systems: challenges and strategies for success O’Kennedy, Richard; Fitzgerald, Jenny; Cassedy, Arabelle Journal of Micromechanics and Microengineering, Vol. 28, Issue 6 https://doi.org/10.1088/1361-6439/aab225	journal	March 2018
CD-Based Microfluidics for Primary Care in Extreme Point-of-Care Settings Smith, Suzanne; Mager, Dario; Perebikovsky, Alexandra Micromachines, Vol. 7, Issue 2 https://doi.org/10.3390/mi7020022	journal	January 2016
Measurement of Carcinoembryonic Antigen in Clinical Serum Samples Using a Centrifugal Microfluidic Device Gao, Zhigang; Chen, Zongzheng; Deng, Jiu Micromachines, Vol. 9, Issue 9 https://doi.org/10.3390/mi9090470	journal	September 2018
A Rapid, Sensitive, and Portable Biosensor Assay for the Detection of Botulinum Neurotoxin Serotype A in Complex Food Matrices Tam, Christina; Flannery, Andrew; Cheng, Luisa Toxins, Vol. 10, Issue 11 https://doi.org/10.3390/toxins10110476	journal	November 2018
CD-based microfluidics for primary care in extreme point-of-care settings Smith, S.; Mager, D.; Perebikovsky, A. Karlsruhe https://doi.org/10.5445/ir/1000053234	text	January 2016

Similar Records

A Portable Immunoassay Platform for Multiplexed Detection of Biotoxins in Clinical and Environmental Samples

Journal Article · Wed Dec 31 23:00:00 EST 2014 · Sandia journal manuscript; Not yet accepted for publication · OSTI ID:1427244

Centrifugal sedimentation immunoassays for multiplexed detection of enteric bacteria in ground water

Journal Article · Thu Dec 31 19:00:00 EST 2015 · Biomicrofluidics · OSTI ID:1427213

Rapid, Sensitive Detection of Botulinum Toxin on a Flexible Microfluidics Platform

Conference · Sat Oct 30 00:00:00 EDT 2004 · OSTI ID:15020037

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Centrifugal microfluidic platform for ultrasensitive detection of botulinum toxin

Citation Formats

References (53)

Cited By (16)

Similar Records

Related Subjects