FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Toxins (Basel) / Rapid Microfluidic Assay for the Detection of Botulinum Neur...
Toxins2016; 8(1); 13; doi: 10.3390/toxins8010013

Rapid Microfluidic Assay for the Detection of Botulinum Neurotoxin in Animal Sera.

Abstract: Potent Botulinum neurotoxins (BoNTs) represent a threat to public health and safety. Botulism is a disease caused by BoNT intoxication that results in muscle paralysis that can be fatal. Sensitive assays capable of detecting BoNTs from different substrates and settings are essential to limit foodborne contamination and morbidity. In this report, we describe a rapid 96-well microfluidic double sandwich immunoassay for the sensitive detection of BoNT-A from animal sera. This BoNT microfluidic assay requires only 5 μL of serum, provides results in 75 min using a standard fluorescence microplate reader and generates minimal hazardous waste. The assay has a <30 pg·mL(-1) limit of detection (LOD) of BoNT-A from spiked human serum. This sensitive microfluidic BoNT-A assay offers a fast and simplified workflow suitable for the detection of BoNT-A from serum samples of limited volume in most laboratory settings.
Get Full Text
Publication Date: 2016-01-04 PubMed ID: 26742073PubMed Central: PMC4728535DOI: 10.3390/toxins8010013Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article is about a method for detecting a dangerous toxin using a rapid, minimal-waste technique that requires only a small sample and basic laboratory equipment.

Research Overview

  • The main focus of the research is to detect Botulinum neurotoxins (BoNTs), substances that pose a serious threat to public health. The researchers devised a rapid, microfluidic test that can detect this toxin in animal sera (blood serum).
  • BoNTs cause botulism, a disease that results in muscle paralysis and can be fatal if not treated. Timely detection of these toxins is crucial to prevent food contamination and reduce the incidence of disease.

Methodology

  • The innovative microfluidic assay for BoNTs that the researchers developed is carried out in a 96-well format and is based on a double sandwich immunoassay.
  • A striking feature of the assay is its sensitivity. It requires only 5 μL of serum, a tiny volume, and the assay is quick, providing results in roughly 75 minutes.
  • In addition to being rapid and sensitive, the assay generates minimal hazardous waste, making it a safer and more sustainable choice than traditional methods for toxin detection.

Results

  • The researchers’ sterilization methods yielded successful results, with the assay capable of detecting less than 30 pg·mL(-1) of BoNT-A in spiked human serum.
  • This demonstrates the test’s sensitivity and suggests it could be used for early detection of BoNT contamination, potentially preventing outbreaks of botulism.

Implications

  • This rapid, microfluidic assay could greatly facilitate the detection of BoNTs, thanks to its speed, efficiency, and minimal waste generation.
  • It has a simple workflow, making it suitable for most laboratory environments, and is capable of analyzing samples of limited volume.
  • Given these advantages, the microfluidic assay could considerably improve public health safeguarding measures by providing a rapid, efficient method for detecting a potentially fatal toxin.

Cite This Article

APA
Babrak L, Lin A, Stanker LH, McGarvey J, Hnasko R. (2016). Rapid Microfluidic Assay for the Detection of Botulinum Neurotoxin in Animal Sera. Toxins (Basel), 8(1), 13. https://doi.org/10.3390/toxins8010013

Publication

Toxins
ISSN: 2072-6651
NlmUniqueID: 101530765
Country: Switzerland
Language: English
Volume: 8
Issue: 1
PII: 13

Researcher Affiliations

Babrak, Lmar
  • Produce Safety & Microbiology Research Unit, United States Department of Agriculture, Agricultural Research Service, 800 Buchanan St, Albany, CA 94710, USA. lmar.babrak@ars.usda.gov.
Lin, Alice
  • Produce Safety & Microbiology Research Unit, United States Department of Agriculture, Agricultural Research Service, 800 Buchanan St, Albany, CA 94710, USA. alice.lin@ars.usda.gov.
Stanker, Larry H
  • Foodborne Toxin Detection and Prevention Unit, United States Department of Agriculture, Agricultural Research Service, 800 Buchanan St, Albany, CA 94710, USA. larry.stanker@ars.usda.gov.
McGarvey, Jeffery
  • Foodborne Toxin Detection and Prevention Unit, United States Department of Agriculture, Agricultural Research Service, 800 Buchanan St, Albany, CA 94710, USA. jeffery.mcgarvey@ars.usda.gov.
Hnasko, Robert
  • Produce Safety & Microbiology Research Unit, United States Department of Agriculture, Agricultural Research Service, 800 Buchanan St, Albany, CA 94710, USA. robert.hnasko@ars.usda.gov.

MeSH Terms

  • Animals
  • Antibodies, Immobilized / immunology
  • Botulinum Toxins, Type A / analysis
  • Botulinum Toxins, Type A / blood
  • Botulinum Toxins, Type A / immunology
  • Cattle
  • Enzyme-Linked Immunosorbent Assay
  • Fabaceae
  • Food, Preserved / analysis
  • Fruit and Vegetable Juices / analysis
  • Horses
  • Humans
  • Limit of Detection
  • Mice
  • Microfluidic Analytical Techniques
  • Neurotoxins / analysis
  • Neurotoxins / blood
  • Neurotoxins / immunology
  • Serum / chemistry
  • Sheep

References

This article includes 24 references
  1. Sobel J. Botulism.. Clin Infect Dis 2005 Oct 15;41(8):1167-73.
    doi: 10.1086/444507pubmed: 16163636google scholar: lookup
  2. Tighe AP, Schiavo G. Botulinum neurotoxins: mechanism of action.. Toxicon 2013 Jun 1;67:87-93.
    doi: 10.1016/j.toxicon.2012.11.011pubmed: 23201505google scholar: lookup
  3. Dembek ZF, Smith LA, Rusnak JM. Botulism: cause, effects, diagnosis, clinical and laboratory identification, and treatment modalities.. Disaster Med Public Health Prep 2007 Nov;1(2):122-34.
    doi: 10.1097/DMP.0b013e318158c5fdpubmed: 18388640google scholar: lookup
  4. Rossetto O, Pirazzini M, Montecucco C. Botulinum neurotoxins: genetic, structural and mechanistic insights.. Nat Rev Microbiol 2014 Aug;12(8):535-49.
    doi: 10.1038/nrmicro3295pubmed: 24975322google scholar: lookup
  5. Burnett JC, Henchal EA, Schmaljohn AL, Bavari S. The evolving field of biodefence: therapeutic developments and diagnostics.. Nat Rev Drug Discov 2005 Apr;4(4):281-97.
    doi: 10.1038/nrd1694pmc: PMC7096857pubmed: 15803193google scholar: lookup
  6. Anniballi F, Fiore A, Löfström C, Skarin H, Auricchio B, Woudstra C, Bano L, Segerman B, Koene M, Båverud V, Hansen T, Fach P, Tevell Aberg A, Hedeland M, Olsson Engvall E, De Medici D. Management of animal botulism outbreaks: from clinical suspicion to practical countermeasures to prevent or minimize outbreaks.. Biosecur Bioterror 2013 Sep;11 Suppl 1:S191-9.
    doi: 10.1089/bsp.2012.0089pubmed: 23971806google scholar: lookup
  7. Ferreira JL. Comparison of amplified ELISA and mouse bioassay procedures for determination of botulinal toxins A, B, E, and F.. J AOAC Int 2001 Jan-Feb;84(1):85-8.
    pubmed: 11234855
  8. Cai S, Singh BR, Sharma S. Botulism diagnostics: from clinical symptoms to in vitro assays.. Crit Rev Microbiol 2007 Apr-Jun;33(2):109-25.
    doi: 10.1080/10408410701364562pubmed: 17558660google scholar: lookup
  9. Ching KH, Lin A, McGarvey JA, Stanker LH, Hnasko R. Rapid and selective detection of botulinum neurotoxin serotype-A and -B with a single immunochromatographic test strip.. J Immunol Methods 2012 Jun 29;380(1-2):23-9.
    doi: 10.1016/j.jim.2012.03.008pubmed: 22504369google scholar: lookup
  10. Cheng LW, Stanker LH. Detection of botulinum neurotoxin serotypes A and B using a chemiluminescent versus electrochemiluminescent immunoassay in food and serum.. J Agric Food Chem 2013 Jan 23;61(3):755-60.
    doi: 10.1021/jf3041963pmc: PMC3598631pubmed: 23265581google scholar: lookup
  11. Singh A, Datta S, Sachdeva A, Maslanka S, Dykes J, Skinner G, Burr D, Whiting RC, Sharma SK. Evaluation of an enzyme-linked immunosorbent assay (ELISA) kit for the detection of botulinum neurotoxins A, B, E, and F in selected food matrices.. Health Secur 2015 Jan-Feb;13(1):37-44.
    pubmed: 25812427doi: 10.1089/hs.2014.0075google scholar: lookup
  12. Singh AK, Stanker LH, Sharma SK. Botulinum neurotoxin: where are we with detection technologies?. Crit Rev Microbiol 2013 Feb;39(1):43-56.
    doi: 10.3109/1040841X.2012.691457pubmed: 22676403google scholar: lookup
  13. Stanker LH, Merrill P, Scotcher MC, Cheng LW. Development and partial characterization of high-affinity monoclonal antibodies for botulinum toxin type A and their use in analysis of milk by sandwich ELISA.. J Immunol Methods 2008 Jul 20;336(1):1-8.
    doi: 10.1016/j.jim.2008.03.003pubmed: 18452945google scholar: lookup
  14. Butler JE. Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays.. Methods 2000 Sep;22(1):4-23.
    doi: 10.1006/meth.2000.1031pubmed: 11020313google scholar: lookup
  15. Bompiani KM, Dickerson TJ. High-throughput screening technologies for botulinum neurotoxins.. Curr Top Med Chem 2014;14(18):2062-80.
    doi: 10.2174/1568026614666141022093857pubmed: 25335886google scholar: lookup
  16. Rosow LK, Strober JB. Infant botulism: review and clinical update.. Pediatr Neurol 2015 May;52(5):487-92.
    doi: 10.1016/j.pediatrneurol.2015.01.006pubmed: 25882077google scholar: lookup
  17. Arnon SS, Schechter R, Inglesby TV, Henderson DA, Bartlett JG, Ascher MS, Eitzen E, Fine AD, Hauer J, Layton M, Lillibridge S, Osterholm MT, O'Toole T, Parker G, Perl TM, Russell PK, Swerdlow DL, Tonat K. Botulinum toxin as a biological weapon: medical and public health management.. JAMA 2001 Feb 28;285(8):1059-70.
    doi: 10.1001/jama.285.8.1059pubmed: 11209178google scholar: lookup
  18. Whitesides GM. The origins and the future of microfluidics.. Nature 2006 Jul 27;442(7101):368-73.
    doi: 10.1038/nature05058pubmed: 16871203google scholar: lookup
  19. Baker CA, Duong CT, Grimley A, Roper MG. Recent advances in microfluidic detection systems.. Bioanalysis 2009 Aug;1(5):967-75.
    doi: 10.4155/bio.09.86pmc: PMC2856342pubmed: 20414455google scholar: lookup
  20. Sanjay ST, Fu G, Dou M, Xu F, Liu R, Qi H, Li X. Biomarker detection for disease diagnosis using cost-effective microfluidic platforms.. Analyst 2015 Nov 7;140(21):7062-81.
    doi: 10.1039/C5AN00780Apmc: PMC4604043pubmed: 26171467google scholar: lookup
  21. Jung W, Han J, Choi J, Ahn C. Point-of-care testing (POCT) diagnostic systems using microfluidic lab-on-a-chip technologies.. Microelectron. Eng. 2015;132:46–57.
    doi: 10.1016/j.mee.2014.09.024google scholar: lookup
  22. Zhang Z, Yu L, Xu L, Hu X, Li P, Zhang Q, Ding X, Feng X. Biotoxin sensing in food and environment via microchip.. Electrophoresis 2014 Jun;35(11):1547-59.
    doi: 10.1002/elps.201300570pubmed: 24723235google scholar: lookup
  23. Scotcher MC, Johnson EA, Stanker LH. Characterization of the epitope region of F1-2 and F1-5, two monoclonal antibodies to Botulinum neurotoxin type A.. Hybridoma (Larchmt) 2009 Oct;28(5):315-25.
    doi: 10.1089/hyb.2009.0022pubmed: 19857112google scholar: lookup
  24. Scotcher MC, McGarvey JA, Johnson EA, Stanker LH. Epitope characterization and variable region sequence of f1-40, a high-affinity monoclonal antibody to botulinum neurotoxin type a (Hall strain).. PLoS One 2009;4(3):e4924.
    doi: 10.1371/journal.pone.0004924pmc: PMC2654115pubmed: 19290051google scholar: lookup

Citations

This article has been cited 7 times.
  1. Rasetti-Escargueil C, Popoff MR. Recent Developments in Botulinum Neurotoxins Detection.. Microorganisms 2022 May 10;10(5).
    doi: 10.3390/microorganisms10051001pubmed: 35630444google scholar: lookup
  2. Chen Y, Li H, Yang L, Wang L, Sun R, Shearer JES, Sun F. Rapid Detection of Clostridium botulinum in Food Using Loop-Mediated Isothermal Amplification (LAMP).. Int J Environ Res Public Health 2021 Apr 21;18(9).
    doi: 10.3390/ijerph18094401pubmed: 33919101google scholar: lookup
  3. Rasetti-Escargueil C, Lemichez E, Popoff MR. Toxemia in Human Naturally Acquired Botulism.. Toxins (Basel) 2020 Nov 13;12(11).
    doi: 10.3390/toxins12110716pubmed: 33202855google scholar: lookup
  4. Lonati D, Schicchi A, Crevani M, Buscaglia E, Scaravaggi G, Maida F, Cirronis M, Petrolini VM, Locatelli CA. Foodborne Botulism: Clinical Diagnosis and Medical Treatment.. Toxins (Basel) 2020 Aug 7;12(8).
    doi: 10.3390/toxins12080509pubmed: 32784744google scholar: lookup
  5. Lee NH, Nahm SH, Choi IS. Real-Time Monitoring of a Botulinum Neurotoxin Using All-Carbon Nanotube-Based Field-Effect Transistor Devices.. Sensors (Basel) 2018 Dec 3;18(12).
    doi: 10.3390/s18124235pubmed: 30513867google scholar: lookup
  6. Tam CC, Flannery AR, Cheng LW. A Rapid, Sensitive, and Portable Biosensor Assay for the Detection of Botulinum Neurotoxin Serotype A in Complex Food Matrices.. Toxins (Basel) 2018 Nov 15;10(11).
    doi: 10.3390/toxins10110476pubmed: 30445734google scholar: lookup
  7. Thirunavukkarasu N, Johnson E, Pillai S, Hodge D, Stanker L, Wentz T, Singh B, Venkateswaran K, McNutt P, Adler M, Brown E, Hammack T, Burr D, Sharma S. Botulinum Neurotoxin Detection Methods for Public Health Response and Surveillance.. Front Bioeng Biotechnol 2018;6:80.
    doi: 10.3389/fbioe.2018.00080pubmed: 29988463google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.