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Home / Equine Research Bank / Sensors (Basel) / Development of Electrochemical DNA Biosensor for Equine Hind...
Sensors (Basel, Switzerland)2021; 21(7); 2319; doi: 10.3390/s21072319

Development of Electrochemical DNA Biosensor for Equine Hindgut Acidosis Detection.

Abstract: The pH drop in the hindgut of the horse is caused by lactic acid-producing bacteria which are abundant when a horse's feeding regime is excessively carbohydrate rich. This drop in pH below six causes hindgut acidosis and may lead to laminitis. Lactic acid-producing bacteria and have been found to produce high amounts of L-lactate and D-lactate, respectively. Early detection of increased levels of these bacteria could allow the horse owner to tailor the horse's diet to avoid hindgut acidosis and subsequent laminitis. Therefore, 16s ribosomal ribonucleic acid (rRNA) sequences were identified and modified to obtain target single stranded deoxyribonucleic acid (DNA) from these bacteria. Complementary single stranded DNAs were designed from the modified target sequences to form capture probes. Binding between capture probe and target single stranded deoxyribonucleic acid (ssDNA) in solution has been studied by gel electrophoresis. Among pairs of different capture probes and target single stranded DNA, hybridization of capture probe 1 (SECP1) and target 1 (SET1) was portrayed as gel electrophoresis. Adsorptive stripping voltammetry was utilized to study the binding of thiol modified SECP1 over gold on glass substrates and these studies showed a consistent binding signal of thiol modified SECP1 and their hybridization with SET1 over the gold working electrode. Cyclic voltammetry and electrochemical impedance spectroscopy were employed to examine the binding of thiol modified SECP1 on the gold working electrode and hybridization of thiol modified SECP1 with the target single stranded DNA. Both demonstrated the gold working electrode surface was modified with a capture probe layer and hybridization of the thiol bound ssDNA probe with target DNA was indicated. Therefore, the proposed electrochemical biosensor has the potential to be used for the detection of the non-synthetic bacterial DNA target responsible for equine hindgut acidosis.
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Publication Date: 2021-03-26 PubMed ID: 33810389PubMed Central: PMC8037926DOI: 10.3390/s21072319Google Scholar: Lookup
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  • 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.

This research outlines the development of an electrochemical biosensor that can detect changes in the bacterial content of a horse’s hindgut, associated with high carbohydrate diets, that might warn of potentially dangerous acidosis and laminitis.

Overview of Study and Objective

  • The study focuses on developing a method to early detect a serious horse health issue – hindgut acidosis – which can lead to a painful and sometimes fatal condition known as laminitis. The cause of this issue often stems from a diet excessively rich in carbohydrates that encourages the growth of lactic acid-producing bacteria.
  • The main purpose of this research is to establish an electrochemical biosensor capable of pinpointing significant increases of lactic acid-producing bacteria in a horse’s hindgut.

Process of Development

  • The first step involved the identification and modification of 16s ribosomal ribonucleic acid (rRNA) sequences to extract target single-stranded deoxyribonucleic acid (DNA) from the lactic acid-producing bacteria.
  • Following this, complementary single-stranded DNAs were designed from these modified target sequences to form capture probes. The binding interaction between these capture probes and target single-stranded (ss) DNA in a solution was then explored using gel electrophoresis. Among different pairings of capture probes and target ssDNA, the bonding of capture probe 1 (SECP1) and target 1 (SET1) was represented through gel electrophoresis.

Methods Used

  • The researchers used Adsorptive Stripping Voltammetry to study the binding pattern of a thiol-modified version of SECP1 over gold on glass substrates. This study revealed a consistent binding signal between the thiol-modified SECP1 and its hybridization with SET1 over a gold working electrode.
  • To further investigate this bonding, they used both cyclic voltammetry and electrochemical impedance spectroscopy. Results from these methods showed that the gold working electrode surface successfully got modified with a capture probe layer and that the ssDNA probe bound with thiol successfully hybridized with the target DNA.

Significance of Findings

  • The results of the research validate the potential of the proposed electrochemical biosensor as a potential tool for the detection of the natural bacterial DNA target whose levels can indicate the risk of equine hindgut acidosis, thus enabling timely dietary intervention.

Cite This Article

APA
Davies J, Thomas C, Rizwan M, Gwenin C. (2021). Development of Electrochemical DNA Biosensor for Equine Hindgut Acidosis Detection. Sensors (Basel), 21(7), 2319. https://doi.org/10.3390/s21072319

Publication

Sensors (Basel, Switzerland)
ISSN: 1424-8220
NlmUniqueID: 101204366
Country: Switzerland
Language: English
Volume: 21
Issue: 7
PII: 2319

Researcher Affiliations

Davies, Joshua
  • School of Natural Sciences, Bangor University, Bangor, Gwynedd, Wales LL57 2UW, UK.
Thomas, Carol
  • School of Natural Sciences, Bangor University, Bangor, Gwynedd, Wales LL57 2UW, UK.
Rizwan, Mohammad
  • School of Natural Sciences, Bangor University, Bangor, Gwynedd, Wales LL57 2UW, UK.
Gwenin, Christopher
  • School of Natural Sciences, Bangor University, Bangor, Gwynedd, Wales LL57 2UW, UK.
  • Department of Chemistry, Xi'an Jiaotong-Liverpool University, 111 Ren'ai Road, Suzhou Industrial Park, Suzhou 215123, China.

MeSH Terms

  • Acidosis
  • Animals
  • Biosensing Techniques
  • DNA
  • DNA Probes
  • Electrochemical Techniques
  • Electrodes
  • Firmicutes
  • Gold
  • Horses
  • Nucleic Acid Hybridization
  • Streptococcus bovis

Grant Funding

  • N/A / Knowledge Economy Skills Scholarship 2 fund
  • N/A / Celtic Advanced Life Sciences Network (CALIN) which is supported by the European Regional Development Fund through the Ireland Wales Cooperation programme

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

References

This article includes 51 references
  1. Milinovich GJ, Trott DJ, Burrell PC, van Eps AW, Thoefner MB, Blackall LL, Al Jassim RA, Morton JM, Pollitt CC. Changes in equine hindgut bacterial populations during oligofructose-induced laminitis.. Environ Microbiol 2006 May;8(5):885-98.
    doi: 10.1111/j.1462-2920.2005.00975.xpubmed: 16623745google scholar: lookup
  2. Milinovich GJ, Trott DJ, Burrell PC, Croser EL, Al Jassim RA, Morton JM, van Eps AW, Pollitt CC. Fluorescence in situ hybridization analysis of hindgut bacteria associated with the development of equine laminitis.. Environ Microbiol 2007 Aug;9(8):2090-100.
    doi: 10.1111/j.1462-2920.2007.01327.xpubmed: 17635552google scholar: lookup
  3. Lecchi C, Dalla Costa E, Lebelt D, Ferrante V, Canali E, Ceciliani F, Stucke D, Minero M. Circulating miR-23b-3p, miR-145-5p and miR-200b-3p are potential biomarkers to monitor acute pain associated with laminitis in horses.. Animal 2018 Feb;12(2):366-375.
    doi: 10.1017/S1751731117001525pubmed: 28689512google scholar: lookup
  4. Nocek JE. Bovine acidosis: implications on laminitis.. J Dairy Sci 1997 May;80(5):1005-28.
    doi: 10.3168/jds.S0022-0302(97)76026-0pubmed: 9178142google scholar: lookup
  5. Van den Berg M, Hoskin SO, Rogers CW, Grinberg A. Fecal pH and Microbial Populations in Thoroughbred Horses During Transition from Pasture to Concentrate Feeding. J. Equine Vet. Sci. 2013;33:215–222.
    doi: 10.1016/j.jevs.2012.06.004google scholar: lookup
  6. Dougal K, de la Fuente G, Harris PA, Girdwood SE, Pinloche E, Newbold CJ. Identification of a core bacterial community within the large intestine of the horse.. PLoS One 2013;8(10):e77660.
    doi: 10.1371/journal.pone.0077660pmc: PMC3812009pubmed: 24204908google scholar: lookup
  7. Al Jassim RA, Scott PT, Trebbin AL, Trott D, Pollitt CC. The genetic diversity of lactic acid producing bacteria in the equine gastrointestinal tract.. FEMS Microbiol Lett 2005 Jul 1;248(1):75-81.
    doi: 10.1016/j.femsle.2005.05.023pubmed: 15953698google scholar: lookup
  8. Marazuela D, Moreno-Bondi MC. Fiber-optic biosensors--an overview.. Anal Bioanal Chem 2002 Mar;372(5-6):664-82.
    pubmed: 11941437doi: 10.1007/s00216-002-1235-9google scholar: lookup
  9. Odenthal KJ, Gooding JJ. An introduction to electrochemical DNA biosensors.. Analyst 2007 Jul;132(7):603-10.
    doi: 10.1039/b701816apubmed: 17592577google scholar: lookup
  10. Drummond TG, Hill MG, Barton JK. Electrochemical DNA sensors.. Nat Biotechnol 2003 Oct;21(10):1192-9.
    doi: 10.1038/nbt873pubmed: 14520405google scholar: lookup
  11. Liu A, Wang K, Weng S, Lei Y, Lin L, Chen W, Lin X, Chen Y. Development of electrochemical DNA biosensors. Trac Trends Anal. Chem. 2012;37:101–111.
    doi: 10.1016/j.trac.2012.03.008google scholar: lookup
  12. Wang J. Electrochemical nucleic acid biosensors. Anal. Chim. Acta. 2002;469:63–71.
    doi: 10.1016/S0003-2670(01)01399-Xgoogle scholar: lookup
  13. Palchetti I, Mascini M. Nucleic acid biosensors for environmental pollution monitoring.. Analyst 2008 Jul;133(7):846-54.
    doi: 10.1039/b802920mpubmed: 18575633google scholar: lookup
  14. Xu K, Chen Y, Okhai TA, Snyman LW. Micro optical sensors based on avalanching silicon light-emitting devices monolithically integrated on chips. Opt. Mater. Express. 2019;9:3985–3997.
    doi: 10.1364/OME.9.003985google scholar: lookup
  15. Cai H, Xu Y, He P-G, Fang Y-Z. Indicator Free DNA Hybridization Detection by Impedance Measurement Based on the DNA-Doped Conducting Polymer Film Formed on the Carbon Nanotube Modified Electrode. Electroanalysis 2003;15:1864–1870.
    doi: 10.1002/elan.200302755google scholar: lookup
  16. Halliwell J, Savage AC, Buckley N, Gwenin C. Electrochemical impedance spectroscopy biosensor for detection of active botulinum neurotoxin. Sens. Bio-Sens. Res. 2014;2:12–15.
    doi: 10.1016/j.sbsr.2014.08.002google scholar: lookup
  17. Savage AC, Buckley N, Halliwell J, Gwenin C. Botulinum neurotoxin serotypes detected by electrochemical impedance spectroscopy.. Toxins (Basel) 2015 May 6;7(5):1544-55.
    doi: 10.3390/toxins7051544pmc: PMC4448162pubmed: 25954998google scholar: lookup
  18. Rizwan M, Hazmi M, Lima SA, Ahmed MU. A highly sensitive electrochemical detection of human chorionic gonadotropin on a carbon nano-onions/gold nanoparticles/polyethylene glycol nanocomposite modified glassy carbon electrode. J. Electroanal. Chem. 2019;833:462–470.
    doi: 10.1016/j.jelechem.2018.12.031google scholar: lookup
  19. Rizwan M, Koh D, Booth MA, Ahmed MU. Combining a gold nanoparticle-polyethylene glycol nanocomposite and carbon nanofiber electrodes to develop a highly sensitive salivary secretory immunoglobulin A immunosensor. Sens. Actuators B Chem. 2018;255:557–563.
    doi: 10.1016/j.snb.2017.08.079google scholar: lookup
  20. Rizwan M, Elma S, Lim SA, Ahmed MU. AuNPs/CNOs/SWCNTs/chitosan-nanocomposite modified electrochemical sensor for the label-free detection of carcinoembryonic antigen.. Biosens Bioelectron 2018 Jun 1;107:211-217.
    doi: 10.1016/j.bios.2018.02.037pubmed: 29471282google scholar: lookup
  21. Ahmad S, Nadeem S, Ullah N. Entropy generation and temperature-dependent viscosity in the study of SWCNT–MWCNT hybrid nanofluid. Appl. Nanosci. 2020;10:5107–5119.
    doi: 10.1007/s13204-020-01306-0google scholar: lookup
  22. Park JY, Park SM. DNA hybridization sensors based on electrochemical impedance spectroscopy as a detection tool.. Sensors (Basel) 2009;9(12):9513-32.
    doi: 10.3390/s91209513pmc: PMC3267184pubmed: 22303136google scholar: lookup
  23. Quan Li P, Piper A, Schmueser I, Mount AR, Corrigan DK. Impedimetric measurement of DNA-DNA hybridisation using microelectrodes with different radii for detection of methicillin resistant Staphylococcus aureus (MRSA).. Analyst 2017 May 30;142(11):1946-1952.
    doi: 10.1039/C7AN90048Apubmed: 28492640google scholar: lookup
  24. Kafka J, Pänke O, Abendroth B, Lisdat F. A label-free DNA sensor based on impedance spectroscopy. Electrochim. Acta. 2008;53:7467–7474.
    doi: 10.1016/j.electacta.2008.01.031google scholar: lookup
  25. Gooding JJ. Electrochemical DNA Hybridization Biosensors. Electroanalysis 2002;14:1149–1156.
    doi: 10.1002/1521-4109(200209)14:17<1149::AID-ELAN1149>3.0.CO;2-8google scholar: lookup
  26. Yang W, Gooding JJ, Hibbert DB. Characterisation of gold electrodes modified with self-assembled monolayers of L-cysteine for the adsorptive stripping analysis of copper. J. Electroanal. Chem. 2001;516:10–16.
    doi: 10.1016/S0022-0728(01)00649-0google scholar: lookup
  27. Shin T, Kim K-N, Lee C-W, Shin SK, Kang H. Self-Assembled Monolayer of L-Cysteine on Au(111): Hydrogen Exchange between Zwitterionic L-Cysteine and Physisorbed Water. J. Phys. Chem. B. 2003;107:11674–11681.
    doi: 10.1021/jp030314jgoogle scholar: lookup
  28. Butterworth A, Blues E, Williamson P, Cardona M, Gray L, Corrigan DK. SAM Composition and Electrode Roughness Affect Performance of a DNA Biosensor for Antibiotic Resistance.. Biosensors (Basel) 2019 Feb 7;9(1).
    doi: 10.3390/bios9010022pmc: PMC6468421pubmed: 30736460google scholar: lookup
  29. Tachibana M, Yoshizawa K, Ogawa A, Fujimoto H, Hoffmann R. Sulfur-gold orbital interactions which determine the structure of alkanethiolate/Au(111) self-assembled monolayer systems. J. Phys. Chem. B. 2002;106:12727–12736.
    doi: 10.1021/jp020993igoogle scholar: lookup
  30. Levicky R, Herne TM, Tarlov MJ, Satija SK. Using self-assembly to control the structure of DNA monolayers on gold: A neutron reflectivity study. J. Am. Chem. Soc. 1998;120:9787–9792.
    doi: 10.1021/ja981897rgoogle scholar: lookup
  31. Merchant B. Gold, the noble metal and the paradoxes of its toxicology.. Biologicals 1998 Mar;26(1):49-59.
    doi: 10.1006/biol.1997.0123pubmed: 9637749google scholar: lookup
  32. Loy JD, Leger L, Workman AM, Clawson ML, Bulut E, Wang B. Development of a multiplex real-time PCR assay using two thermocycling platforms for detection of major bacterial pathogens associated with bovine respiratory disease complex from clinical samples.. J Vet Diagn Invest 2018 Nov;30(6):837-847.
    doi: 10.1177/1040638718800170pmc: PMC6505850pubmed: 30239324google scholar: lookup
  33. Blanco Vázquez C, Alonso-Hearn M, Juste RA, Canive M, Iglesias T, Iglesias N, Amado J, Vicente F, Balseiro A, Casais R. Detection of latent forms of Mycobacterium avium subsp. paratuberculosis infection using host biomarker-based ELISAs greatly improves paratuberculosis diagnostic sensitivity.. PLoS One 2020;15(9):e0236336.
    doi: 10.1371/journal.pone.0236336pmc: PMC7470414pubmed: 32881863google scholar: lookup
  34. Halliwell J, Gwenin C. A label free colorimetric assay for the detection of active botulinum neurotoxin type A by SNAP-25 conjugated colloidal gold.. Toxins (Basel) 2013 Aug 6;5(8):1381-91.
    doi: 10.3390/toxins5081381pmc: PMC3760041pubmed: 23925142google scholar: lookup
  35. Heeroma AJ, Gwenin C. Development of Solid-Phase RPA on a Lateral Flow Device for the Detection of Pathogens Related to Sepsis.. Sensors (Basel) 2020 Jul 28;20(15).
    doi: 10.3390/s20154182pmc: PMC7436017pubmed: 32731402google scholar: lookup
  36. Peng H-P, Hu Y, Liu P, Deng Y-N, Wang P, Chen W, Liu A-L, Chen Y-Z, Lin X-H. Label-free electrochemical DNA biosensor for rapid detection of mutidrug resistance gene based on Au nanoparticles/toluidine blue–graphene oxide nanocomposites. Sens. Actuators B Chem. 2015;207:269–276.
    doi: 10.1016/j.snb.2014.10.059google scholar: lookup
  37. Johnson BN, Mutharasan R. A cantilever biosensor-based assay for toxin-producing cyanobacteria Microcystis aeruginosa using 16S rRNA.. Environ Sci Technol 2013;47(21):12333-41.
    doi: 10.1021/es402925kpubmed: 24070168google scholar: lookup
  38. Liu C, Zeng GM, Tang L, Zhang Y, Li YP, Liu YY, Li Z, Wu MS, Luo J. Electrochemical detection of Pseudomonas aeruginosa 16S rRNA using a biosensor based on immobilized stem-loop structured probe.. Enzyme Microb Technol 2011 Aug 10;49(3):266-71.
    doi: 10.1016/j.enzmictec.2011.06.011pubmed: 22112510google scholar: lookup
  39. Liu CC, Yeung CY, Chen PH, Yeh MK, Hou SY. Salmonella detection using 16S ribosomal DNA/RNA probe-gold nanoparticles and lateral flow immunoassay.. Food Chem 2013 Dec 1;141(3):2526-32.
    doi: 10.1016/j.foodchem.2013.05.089pubmed: 23870991google scholar: lookup
  40. Amann RI, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations.. Appl Environ Microbiol 1990 Jun;56(6):1919-25.
    doi: 10.1128/AEM.56.6.1919-1925.1990pmc: PMC184531pubmed: 2200342google scholar: lookup
  41. Henihan G, Schulze H, Corrigan DK, Giraud G, Terry JG, Hardie A, Campbell CJ, Walton AJ, Crain J, Pethig R, Templeton KE, Mount AR, Bachmann TT. Label- and amplification-free electrochemical detection of bacterial ribosomal RNA.. Biosens Bioelectron 2016 Jul 15;81:487-494.
    doi: 10.1016/j.bios.2016.03.037pubmed: 27016627google scholar: lookup
  42. Hoy MA. Insert Molecular Genetics. .
  43. Luo C, Tang H, Cheng W, Yan L, Zhang D, Ju H, Ding S. A sensitive electrochemical DNA biosensor for specific detection of Enterobacteriaceae bacteria by Exonuclease III-assisted signal amplification.. Biosens Bioelectron 2013 Oct 15;48:132-7.
    doi: 10.1016/j.bios.2013.03.084pubmed: 23669045google scholar: lookup
  44. Brown TA. Gene Cloning & DNA Analysis. .
  45. Kasten FH. Cytochemical studies with acridine orange and the influence of dye contaminants in the staining of nucleic acids.. Int Rev Cytol 1967;21:141-202.
    pubmed: 5338238doi: 10.1016/s0074-7696(08)60814-1google scholar: lookup
  46. McMaster GK, Carmichael GG. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.. Proc Natl Acad Sci U S A 1977 Nov;74(11):4835-8.
    doi: 10.1073/pnas.74.11.4835pmc: PMC432050pubmed: 73185google scholar: lookup
  47. LERMAN LS. Structural considerations in the interaction of DNA and acridines.. J Mol Biol 1961 Feb;3:18-30.
    doi: 10.1016/S0022-2836(61)80004-1pubmed: 13761054google scholar: lookup
  48. LERMAN LS. The structure of the DNA-acridine complex.. Proc Natl Acad Sci U S A 1963 Jan 15;49(1):94-102.
    doi: 10.1073/pnas.49.1.94pmc: PMC300634pubmed: 13929834google scholar: lookup
  49. Faria HAM, Zucolotto V. Label-free electrochemical DNA biosensor for zika virus identification.. Biosens Bioelectron 2019 Apr 15;131:149-155.
    doi: 10.1016/j.bios.2019.02.018pubmed: 30831416google scholar: lookup
  50. Rant U, Arinaga K, Fujita S, Yokoyama N, Abstreiter G, Tornow M. Structural properties of oligonucleotide monolayers on gold surfaces probed by fluorescence investigations.. Langmuir 2004 Nov 9;20(23):10086-92.
    doi: 10.1021/la0492963pubmed: 15518498google scholar: lookup
  51. Rant U, Arinaga K, Tornow M, Kim YW, Netz RR, Fujita S, Yokoyama N, Abstreiter G. Dissimilar kinetic behavior of electrically manipulated single- and double-stranded DNA tethered to a gold surface.. Biophys J 2006 May 15;90(10):3666-71.
    doi: 10.1529/biophysj.105.078857pmc: PMC1440747pubmed: 16473909google scholar: lookup

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