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Methods in molecular biology (Clifton, N.J.)2024; 2742; 47-67; doi: 10.1007/978-1-0716-3561-2_4

Constructing an ELISA for Detection of Anti-Borrelia in Wildlife and Agricultural Animals.

Abstract: Zoonotic diseases have major impacts on human and animal health, as well as being ecologically significant. Lyme Borreliosis or Lyme disease, caused by infection by pathogenic members of the Borrelia genus, is among these zoonotic diseases. Serology is one of the most accessible means for indirect surveillance of pathogen presence by monitoring the presence, abundance, and type of immune response to the pathogen or pathogen-associated epitopes. Serological surveillance of wild animals is important as wild animals are the primary reservoirs of many zoonotic diseases. Similarly, serological surveillance of agricultural animals is important due to their economic importance, in addition to animal welfare concerns. However, serology in any non-model animal such as wildlife or agricultural animals is difficult because serology necessarily relies on blood samples from the animals being tested. While companion or laboratory animals are generally sufficiently accustomed to humans that blood samples can be obtained, obtaining blood samples from wild or agricultural animals is more challenging. This initial challenge is compounded by the absence of validated serological tools to evaluate antibody titres in the sera. In this chapter, we provide methods for constructing an ELISA for the detection of anti-Borrelia antibodies in non-model animals, using studies on horses and cows as a proof of principle. The methods focus on the problems specific to non-model animals including obtaining sera, options for determining positive and negative controls without the ability to perform controlled infections, and methods for test optimization and validation.
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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Publication Date: 2024-01-02 PubMed ID: 38165614PubMed Central: 5960580DOI: 10.1007/978-1-0716-3561-2_4Google Scholar: Lookup
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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.

Overview

  • This research focuses on developing an ELISA (enzyme-linked immunosorbent assay) to detect antibodies against Borrelia, the bacteria causing Lyme disease, in wildlife and agricultural animals.
  • The study addresses challenges related to serological testing in non-model animals, such as obtaining samples and validating test accuracy without controlled infection experiments.

Introduction to the Research

  • Zoonotic diseases, which can spread from animals to humans, significantly affect health and ecosystems.
  • Lyme disease, caused by Borrelia bacteria, is one such zoonotic disease with important public health implications.
  • Monitoring the presence of these pathogens is essential for disease surveillance and control.
  • Serology, the study of blood serum to detect immune responses, is a key surveillance tool because it can identify past or current infections indirectly by detecting antibodies.
  • Wild animals often serve as reservoirs for zoonoses, making their serological surveillance crucial.
  • Agricultural animals also require serological monitoring due to economic impacts and animal health concerns.

Challenges in Serological Surveillance of Non-model Animals

  • Unlike laboratory or companion animals, wildlife and agricultural animals are not as easily handled for blood sampling.
  • The physical challenge of blood collection is the first barrier for performing serology on these animals.
  • There is a scarcity of validated serological tests tailored to non-model animals.
  • Without well-established reference standards or the ability to perform controlled infections, determining accurate positive and negative controls for the test is difficult.
  • This lack complicates the standardization, optimization, and validation of assays such as ELISA in non-model species.

Methodological Approach of the Study

  • The study proposes a practical framework for constructing an ELISA to detect anti-Borrelia antibodies in non-model animals.
  • Horses and cows are used as example species to demonstrate applicability, reflecting both wildlife-adjacent and agricultural contexts.
  • Key components of the methodology include:
    • Careful procedures to obtain blood sera from non-model animals while minimizing animal distress and logistical difficulty.
    • Strategies to identify or approximate positive and negative antibody controls without performing experimental infections, such as using known naturally infected samples or closely related species’ sera.
    • Test optimization techniques aimed at improving ELISA sensitivity and specificity tailored to these species’ antibody profiles.
    • Validation approaches ensuring the ELISA results are reliable despite the lack of standard reference sera or controlled infection data.

Significance and Impact

  • This work contributes an important tool for expanding zoonotic disease surveillance beyond traditional model species.
  • Developing validated ELISAs for non-model animals facilitates monitoring Borrelia exposure in wildlife and agriculturally important animals.
  • Broader use of these methods can improve early detection of Lyme disease reservoirs and inform public health and veterinary interventions.
  • It also addresses a notable gap in zoonotic disease surveillance capabilities by adapting serological methodologies to less-studied animal populations.

Cite This Article

APA
Bland J, McGowan C, Bush E, Lloyd V. (2024). Constructing an ELISA for Detection of Anti-Borrelia in Wildlife and Agricultural Animals. Methods Mol Biol, 2742, 47-67. https://doi.org/10.1007/978-1-0716-3561-2_4

Publication

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
NlmUniqueID: 9214969
Country: United States
Language: English
Volume: 2742
Pages: 47-67

Researcher Affiliations

Bland, Julia
  • Department of Biology, Mount Allison University, Sackville, NB, Canada.
  • Atlantic Veterinary College, Charlottetown, PE, Canada.
McGowan, Caitlin
  • Atlantic Veterinary College, Charlottetown, PE, Canada.
  • Nova Scotia, Society for Prevention of Cruelty to Animals (SPCA), Dartmouth, NS, Canada.
Bush, Emma
  • Department of Biology, Mount Allison University, Sackville, NB, Canada.
  • Atlantic Veterinary College, Charlottetown, PE, Canada.
Lloyd, Vett
  • Department of Biology, Mount Allison University, Sackville, NB, Canada. vlloyd@mta.ca.

MeSH Terms

  • Female
  • Animals
  • Humans
  • Horses
  • Cattle
  • Borrelia
  • Animals, Wild
  • Lyme Disease / diagnosis
  • Lyme Disease / veterinary
  • Zoonoses
  • Enzyme-Linked Immunosorbent Assay / methods
  • Antibodies, Bacterial

References

This article includes 53 references
  1. Jones KE, Patel NG, Levy MA. Global trends in emerging infectious diseases. Nature 451(7181):990–993.
    doi: 10.1038/nature06536pubmed: 18288193pmc: 5960580google scholar: lookup
  2. Dong Y, Zhou G, Cao W. Global seroprevalence and sociodemographic characteristics of Borrelia burgdorferi sensu lato in human populations: a systematic review and meta-analysis. BMJ Glob Health 7(6):e007744.
    doi: 10.1136/bmjgh-2021-007744pubmed: 35697507pmc: 9185477google scholar: lookup
  3. Kučerová HL, Žákovská A, Marková J, Bártová E. Detection of antibodies to Borrelia burgdorferi s.l. in wild small mammals and sensitivity of PCR and cultivation. Vet Microbiol 230:241–243.
    doi: 10.1016/j.vetmic.2019.02.004pubmed: 30827395google scholar: lookup
  4. Zinck CB, Priest JM, Shutler D. Detection of Borrelia spp., Ehrlichia canis, Anaplasma phagocytophilum, and Dirofilaria immitis in Eastern Coyotes (Canis latrans) in Nova Scotia, Canada. J Wildl Dis 57(3):678–682.
    doi: 10.7589/JWD-D-20-00188pubmed: 33956091google scholar: lookup
  5. Burgess EC. Borrelia burgdorferi infection in Wisconsin horses and cows. Ann N Y Acad Sci 539:235–243.
    doi: 10.1111/j.1749-6632.1988.tb31857.xpubmed: 3190095google scholar: lookup
  6. Hahn CN, Mayhew IG, Whitwell KE. A possible case of Lyme Borreliosis in a horse in the UK. Equine Vet J 28(1):84–88.
    doi: 10.1111/j.2042-3306.1996.tb01595.x´pubmed: 8565961google scholar: lookup
  7. Burgess B. British Columbia. Lyme disease in horses. Can Vet J 29(4):393-4.7.
  8. Tylewska-Wierzbanowska S, Chmielewski T. Limitation of serological testing for Lyme Borreliosis: evaluation of ELISA and western blot in comparison with PCR and culture methods. Wien Klin Wochenschr 114(13–14):601–605.
  9. Chang YF, Novosol V, McDonough SP. Experimental infection of ponies with Borrelia burgdorferi by exposure to Ixodid ticks. Vet Pathol 37(1):68–76.
    doi: 10.1354/vp.37-1-68pubmed: 10643983google scholar: lookup
  10. Ben Said M, Belkahia H, Alberti A. First molecular evidence of Borrelia burgdorferi sensu lato in goats, sheep, cattle and camels in Tunisia. Ann Agric Environ Med 23(3):442–447.
    doi: 10.5604/12321966.1219184pubmed: 27660865google scholar: lookup
  11. George JE, Pound JM, Davey RB. Chemical control of ticks on cattle and the resistance of these parasites to acaricides. Parasitology 129(Suppl):S353–S366.
    doi: 10.1017/s0031182003004682pubmed: 15938518google scholar: lookup
  12. Heyman P, Cochez C, Hofhuis A. A clear and present danger: tick-borne diseases in Europe. Expert Rev Anti-Infect Ther 8(1):33–50.
    doi: 10.1586/eri.09.118pubmed: 20014900google scholar: lookup
  13. Ji B, Collins MT. Seroepidemiologic survey of Borrelia burgdorferi exposure of dairy cattle in Wisconsin. Am J Vet Res 55(9):1228–1231.
    pubmed: 7802388
  14. Post JE, Shaw EE, Wright SD. Suspected Borreliosis in cattle. Ann N Y Acad Sci 539(1):488–488.
    doi: 10.1111/j.1749-6632.1988.tb31916.xgoogle scholar: lookup
  15. Sharma SP, Amanfu W, Losho TC. Bovine Borreliosis in Botswana. Onderstepoort J Vet Res 67(3):221–223.
    pubmed: 11131124
  16. Stefanciková A, Adaszek Ł, Pet’ko B. Serological evidence of Borrelia burgdorferi sensu lato in horses and cattle from Poland and diagnostic problems of Lyme Borreliosis. Ann Agric Environ Med 15(1):37–43.
  17. Zarkov I, Marinov M. The Lyme disease: results of a serological study in sheep, cows and dogs in Bulgaria. Rev Médecine Vét 154(5):363–366.
  18. Zintl A, Moutailler S, Stuart P. Ticks and Tick-borne diseases in Ireland. Ir Vet J 70:4.
    doi: 10.1186/s13620-017-0084-ypubmed: 28163889pmc: 5282849google scholar: lookup
  19. Lischer CJ, Leutenegger CM, Braun U, Lutz H. Diagnosis of Lyme disease in two cows by the detection of Borrelia burgdorferi DNA. Vet Rec 146(17):497–499.
    doi: 10.1136/vr.146.17.497pubmed: 10887997google scholar: lookup
  20. Parker JL, White KK. Lyme Borreliosis in cattle and horses: a review of the literature. Cornell Vet 82(3):253–274.
    pubmed: 1643876
  21. Blowey RW, Carter SD, White AG, Barnes A. Borrelia burgdorferi infections in UK cattle: a possible association with digital dermatitis. Vet Rec 135(24):577–578.
    pubmed: 7886898
  22. Wagner B, Freer H, Rollins A. Development of a multiplex assay for the detection of antibodies to Borrelia burgdorferi in horses and its validation using Bayesian and conventional statistical methods. Vet Immunol Immunopathol 144(3–4):374–381.
    doi: 10.1016/j.vetimm.2011.08.005pubmed: 21890217google scholar: lookup
  23. Burgess EC, Wachal MD, Cleven TD. Borrelia burgdorferi infection in dairy cows, rodents, and birds from four Wisconsin dairy farms. Vet Microbiol 35(1–2):61–77.
    doi: 10.1016/0378-1135(93)90116-opubmed: 8362496google scholar: lookup
  24. Richter D, Matuschka FR. Modulatory effect of cattle on risk for Lyme disease. Emerg Infect Dis 12(12):1919–1923.
    doi: 10.3201/eid1212.051552pubmed: 17326945pmc: 3291337google scholar: lookup
  25. Richter D, Matuschka FR. Elimination of Lyme disease spirochetes from ticks feeding on domestic ruminants. Appl Environ Microbiol 76(22):7650–7652.
    doi: 10.1128/AEM.01649-10pubmed: 20870789pmc: 2976204google scholar: lookup
  26. Sperling JLH, Sperling FAH. Lyme Borreliosis in Canada: biological diversity and diagnostic complexity from an entomological perspective. Can Entomol 141(6):521–549.
    doi: 10.4039/n08-CPA04google scholar: lookup
  27. Stefancíková A, Stĕpánová G, Derdáková M. Serological evidence for Borrelia burgdorferi infection associated with clinical signs in dairy cattle in Slovakia. Vet Res Commun 26(8):601–611.
    doi: 10.1023/a:1020912618950pubmed: 12507035google scholar: lookup
  28. Divers TJ, Gardner RB, Madigan JE. Borrelia burgdorferi infection and Lyme disease in North American horses: a consensus statement. J Vet Intern Med 32(2):617–632.
    doi: 10.1111/jvim.15042pubmed: 29469222pmc: 5866975google scholar: lookup
  29. Arnold WK, Savage CR, Brissette CA. RNA-Seq of Borrelia burgdorferi in Multiple Phases of Growth Reveals Insights into the Dynamics of Gene Expression, Transcriptome Architecture, and Noncoding RNAs. PLoS One 11(10):e0164165.
    doi: 10.1136/bmj.308.6943.1552pubmed: 8019315pmc: 2540489google scholar: lookup
  30. Altman DG, Bland JM. Diagnostic tests. 1: sensitivity and specificity. BMJ 308(6943):1552.
    doi: 10.1136/bmj.308.6943.1552google scholar: lookup
  31. Carroll JA, Stewart PE, Rosa P. An enhanced GFP reporter system to monitor gene expression in Borrelia burgdorferi. Microbiology (Reading) 149(Pt 7):1819–1828.
    doi: 10.1099/mic.0.26165-0pubmed: 12855733google scholar: lookup
  32. Hyde JA, Trzeciakowski JP, Skare JT. Borrelia burgdorferi alters its gene expression and antigenic profile in response to CO2 levels. J Bacteriol 189(2):437–445.
    doi: 10.1128/JB.01109-06pubmed: 17098904google scholar: lookup
  33. Seshu J, Boylan JA, Gherardini FC, Skare JT. Dissolved oxygen levels alter gene expression and antigen profiles in Borrelia burgdorferi. Infect Immun 72(3):1580–1586.
    doi: 10.1128/IAI.72.3.1580-1586.2004pubmed: 14977964pmc: 356058google scholar: lookup
  34. Tokarz R, Anderton JM, Katona LI, Benach JL. Combined effects of blood and temperature shift on Borrelia burgdorferi gene expression as determined by whole genome DNA array. Infect Immun 72(9):5419–5432.
    doi: 10.1128/IAI.72.9.5419-5432.2004pubmed: 15322040pmc: 517457google scholar: lookup
  35. Brorson Ø, Brorson SH, Scythes J. Destruction of spirochete Borrelia burgdorferi round-body propagules (RBs) by the antibiotic tigecycline. Proc Natl Acad Sci U S A 106(44):18656–18661.
    doi: 10.1073/pnas.0908236106pubmed: 19843691pmc: 2774030google scholar: lookup
  36. Feng J, Shi W, Zhang S. A drug combination screen identifies drugs active against amoxicillin-induced round bodies of in vitro Borrelia burgdorferi persisters from an FDA drug library. Front Microbiol 7:743.
    doi: 10.3389/fmicb.2016.00743pubmed: 27242757pmc: 4876775google scholar: lookup
  37. Feng J, Li T, Yee R. Stationary phase persister/biofilm microcolony of Borrelia burgdorferi causes more severe disease in a mouse model of Lyme arthritis: implications for understanding persistence, Post-treatment Lyme Disease Syndrome (PTLDS), and treatment failure. Discov Med 27(148):125–138.
    pubmed: 30946803
  38. Meriläinen L, Brander H, Herranen A. Pleomorphic forms of Borrelia burgdorferi induce distinct immune responses. Microbes Infect 18(7–8):484–495.
    doi: 10.1016/j.micinf.2016.04.002pubmed: 27139815google scholar: lookup
  39. Rudenko N, Golovchenko M, Kybicova K, Vancova M. Metamorphoses of Lyme disease spirochetes: phenomenon of Borrelia persisters. Parasit Vectors 12(1):237.
    doi: 10.1186/s13071-019-3495-7pubmed: 31097026pmc: 6521364google scholar: lookup
  40. Frey A, Di Canzio J, Zurakowski D. A statistically defined endpoint titer determination method for immunoassays. J Immunol Methods 221(1–2):35–41.
    doi: 10.1016/s0022-1759(98)00170-7pubmed: 9894896google scholar: lookup
  41. Munro HJ, Ogden NH, Mechai S. Genetic diversity of Borrelia garinii from Ixodes uriae collected in seabird colonies of the northwestern Atlantic Ocean. Ticks Tick Borne Dis 10(6):101255.
    doi: 10.1016/j.ttbdis.2019.06.014pubmed: 31280947google scholar: lookup
  42. Ogden NH, Barker IK, Francis CM. How far north are migrant birds transporting the tick Ixodes scapularis in Canada? Insights from stable hydrogen isotope analyses of feathers. Ticks Tick Borne Dis 6(6):715–720.
    doi: 10.1016/j.ttbdis.2015.06.004pubmed: 26100493google scholar: lookup
  43. Scott JD. Studies abound on how far north Ixodes scapularis ticks are transported by birds. Ticks Tick Borne Dis 7(2):327–328.
    doi: 10.1016/j.ttbdis.2015.12.001pubmed: 26739029google scholar: lookup
  44. Scott JD, Fernando K, Banerjee SN. Birds disperse ixodid (Acari: Ixodidae) and Borrelia burgdorferi-infected ticks in Canada. J Med Entomol 8(4):493–500.
    doi: 10.1603/0022-2585-38.4.493google scholar: lookup
  45. Scott JD, Anderson JF, Durden LA. Widespread dispersal of Borrelia burgdorferi-infected ticks collected from songbirds across Canada. J Parasitol 98(1):49–59.
    doi: 10.1645/GE-2874.1pubmed: 21864130google scholar: lookup
  46. Smith RP Jr, Muzaffar SB, Lavers J. Borrelia garinii in seabird ticks (Ixodes uriae), Atlantic Coast, North America.. Emerg Infect Dis 2006 12(12):1909–1912.
    doi: 10.3201/eid1212.060448pubmed: 17326943pmc: 3291351google scholar: lookup
  47. Branda JA, Lemieux JE, Blair L. Detection of Borrelia burgdorferi cell-free DNA in human plasma samples for improved diagnosis of early Lyme Borreliosis.. Clin Infect Dis 2021 73(7):e2355–e2361.
    doi: 10.1093/cid/ciaa858pubmed: 32584965google scholar: lookup
  48. Cerar T, Ruzić-Sabljić E, Glinsek U. Comparison of PCR methods and culture for the detection of Borrelia spp. in patients with erythema migrans.. Clin Microbiol Infect 2008 14(7):653–658.
    doi: 10.1111/j.1469-0691.2008.02013.xpubmed: 18558937google scholar: lookup
  49. Association of Public Health Laboratories. Suggested reporting language, interpretation and guidance regarding Lyme disease serologic test results.. Association of Public Health Laboratories 2021.
  50. Pillai-Kastoori L, Schutz-Geschwender AR, Harford JA. A systematic approach to quantitative Western blot analysis.. Anal Biochem 2020 593:113608.
    doi: 10.1016/j.ab.2020.113608pubmed: 32007473google scholar: lookup
  51. Parikh R, Mathai A, Parikh S. Understanding and using sensitivity, specificity and predictive values.. Indian J Ophthalmol 2008 56(1):45–50.
    doi: 10.4103/0301-4738.37595pubmed: 18158403pmc: 2636062google scholar: lookup
  52. Loss SR, Noden BH, Hamer GL, Hamer SA. A quantitative synthesis of the role of birds in carrying ticks and tick-borne pathogens in North America.. Oecologia 2016 182(4):947–959.
    doi: 10.1007/s00442-016-3731-1pubmed: 27670413google scholar: lookup
  53. Altman DG, Bland JM. Statistics notes: diagnostic tests 2: predictive values.. BMJ 1994 309(6947):102.
    doi: 10.1136/bmj.309.6947.102pubmed: 8038641pmc: 2540558google scholar: lookup

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