Performance of a commercially available in-clinic ELISA for the detection of antibodies against Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi and Dirofilaria immitis antigen in dogs

Ramaswamy ChandrashekarFrom Immunoassay R&D, IDEXX Laboratories, 1 IDEXX Dr, Westbrook, ME 04092

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Celine A. MainvilleFrom Immunoassay R&D, IDEXX Laboratories, 1 IDEXX Dr, Westbrook, ME 04092

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Melissa J. BeallFrom Immunoassay R&D, IDEXX Laboratories, 1 IDEXX Dr, Westbrook, ME 04092

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Thomas O'ConnorFrom Immunoassay R&D, IDEXX Laboratories, 1 IDEXX Dr, Westbrook, ME 04092

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Matthew D. EbertsLakeland Veterinary Hospital, 7372 Woida Rd, Baxter, MN 56425

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A. Rick AllemanDepartment of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32608

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Stephen D. GauntDepartment of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803

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Edward B. BreitschwerdtDepartment of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606

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Abstract

Objective—To evaluate the sensitivity and specificity of a commercially available in-clinic ELISA for detection of heartworm infection and tick-borne diseases in dogs.

Sample Population—846 serum, plasma, or blood samples obtained from dogs.

Procedures—Samples were evaluated via the in-clinic ELISA to detect antibodies against Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi and Dirofilaria immitis (heartworm) antigen. True infection or immunologic status of samples was assessed by use of results of necropsy, an antigen assay for D immitis, and immunofluorescence assay or western blot analysis for antibodies against B burgdorferi, E canis, and A phagocytophilum.

Results—Sensitivity and specificity of the in-clinic ELISA for detection of heartworm antigen (99.2% and 100%, respectively), antibodies against B burgdorferi (98.8% and 100%, respectively), and antibodies against E canis (96.2% and 100%, respectively) were similar to results for a similar commercial ELISA. In samples obtained from dogs in the northeast and upper Midwest of the United States, sensitivity and specificity of the in-clinic ELISA for antibodies against Anaplasma spp were 99.1% and 100%, respectively, compared with results for an immunofluorescence assay. Samples from 2 dogs experimentally infected with the NY18 strain of A phagocytophilum were tested by use of the in-clinic ELISA, and antibodies against A phagocytophilum were detected by 8 days after inoculation. Antibodies against Anaplasma platys in experimentally infected dogs cross-reacted with the A phagocytophilum analyte. Coinfections were identified in several of the canine serum samples.

Conclusions and Clinical Relevance—The commercially available in-clinic ELISA could be used by veterinarians to screen dogs for heartworm infection and for exposure to tick-borne pathogens.

Abstract

Objective—To evaluate the sensitivity and specificity of a commercially available in-clinic ELISA for detection of heartworm infection and tick-borne diseases in dogs.

Sample Population—846 serum, plasma, or blood samples obtained from dogs.

Procedures—Samples were evaluated via the in-clinic ELISA to detect antibodies against Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi and Dirofilaria immitis (heartworm) antigen. True infection or immunologic status of samples was assessed by use of results of necropsy, an antigen assay for D immitis, and immunofluorescence assay or western blot analysis for antibodies against B burgdorferi, E canis, and A phagocytophilum.

Results—Sensitivity and specificity of the in-clinic ELISA for detection of heartworm antigen (99.2% and 100%, respectively), antibodies against B burgdorferi (98.8% and 100%, respectively), and antibodies against E canis (96.2% and 100%, respectively) were similar to results for a similar commercial ELISA. In samples obtained from dogs in the northeast and upper Midwest of the United States, sensitivity and specificity of the in-clinic ELISA for antibodies against Anaplasma spp were 99.1% and 100%, respectively, compared with results for an immunofluorescence assay. Samples from 2 dogs experimentally infected with the NY18 strain of A phagocytophilum were tested by use of the in-clinic ELISA, and antibodies against A phagocytophilum were detected by 8 days after inoculation. Antibodies against Anaplasma platys in experimentally infected dogs cross-reacted with the A phagocytophilum analyte. Coinfections were identified in several of the canine serum samples.

Conclusions and Clinical Relevance—The commercially available in-clinic ELISA could be used by veterinarians to screen dogs for heartworm infection and for exposure to tick-borne pathogens.

Contributor Notes

Presented as a poster at the 25th Annual Forum of the American College of Veterinary Internal Medicine, Seattle, June 2007.

Address correspondence to Dr. Chandrashekar (chandra@idexx.com).
  • 1

    Woldehiwet Z, Scott GR. Tick-borne (pasture) fever. In: Woldehiwet Z, Ristic M, eds. Rickettsial and chlamydial diseases of domestic animals. Oxford, England: Pergamon Press, 1993;209214.

    • Search Google Scholar
    • Export Citation
  • 2

    Chen SM, Dumler JS, Bakken JS, et al. Identification of a granulocytotropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol 1994; 32:589595.

    • Search Google Scholar
    • Export Citation
  • 3

    Alleman RA, Wamsley HL. An update on anaplasmosis in dogs. Vet Med (Praha) 2008; 103:212222.

  • 4

    Nicholson WL, Comer JA, Sumner JW. An indirect immunofluorescence assay using a cell culture-derived antigen for detection of antibodies to the agent of human granulocytic ehrlichiosis. J Clin Microbiol 1997; 35:15101516.

    • Search Google Scholar
    • Export Citation
  • 5

    Suksawat J, Hegarty BC, Breitschwerdt EB. Seroprevalence of Ehrlichia canis, Ehrlichia equi, and Ehrlichia risticii in sick dogs from North Carolina and Virginia. J Vet Intern Med 2000; 14:5055.

    • Search Google Scholar
    • Export Citation
  • 6

    Fritz CL. Emerging tick-borne diseases. Vet Clin North Am Small Anim Pract 2009; 39:265278.

  • 7

    Levy SA. Use of a C6 ELISA test to evaluate the efficacy of a whole-cell bacterin for the prevention of naturally transmitted canine Borrelia burgdorferi infection. Vet Ther 2002; 3:420424.

    • Search Google Scholar
    • Export Citation
  • 8

    Levy SA, O'Connor TP, Hanscom JL, et al. Quantitative measurement of C6 antibody following antibiotic treatment of Borrelia burgdorferi antibody-positive nonclinical dogs. Clin Vaccine Immunol 2008; 15:115119.

    • Search Google Scholar
    • Export Citation
  • 9

    Rikihisa Y. Cross-reacting antigens between Neorickettsia helminthoeca and Ehrlichia species, shown by immunofluorescence and western immunoblotting. J Clin Microbiol 1991; 29:20242029.

    • Search Google Scholar
    • Export Citation
  • 10

    Atkins CE. Comparison of results of three commercial heart-worm antigen test kits in dogs with low heartworm burdens. J Am Vet Med Assoc 2003; 222:12211223.

    • Search Google Scholar
    • Export Citation
  • 11

    Goodman JL, Nelson C, Vitale B, et al. Direct cultivation of the causative agent of human granulocytic ehrlichiosis. N Engl J Med 1996; 334:209215.

    • Search Google Scholar
    • Export Citation
  • 12

    Simpson RM, Gaunt SD, Hair JA, et al. Evaluation of Rhipicephalus sanguineus as a potential biologic vector of Ehrlichia platys. Am J Vet Res 1991; 52:15371541.

    • Search Google Scholar
    • Export Citation
  • 13

    Zhi N, Ohashi N, Tajima T, et al. Transcript heterogeneity of the p44 multigene family in a human granulocytic ehrlichiosis agent transmitted by ticks. Infect Immun 2002; 70:11751184.

    • Search Google Scholar
    • Export Citation
  • 14

    O'Connor T P, Esty E, Machenry P, et al T. Performance evaluation of Ehrlichia canis and Borrelia burgdorferi peptides in a new Dirofilaria immitis combination assay, in Proceedings. Recent Adv Heartworm Dis Symp Am Heartworm Soc 2001;7784.

    • Search Google Scholar
    • Export Citation
  • 15

    Chandrashekar R, Daniluk D, Moffitt S, et al. Serologic diagnosis of equine borreliosis: evaluation of an in-clinic enzyme-linked immunosorbent assay (SNAP® 4Dx®). Int J Appl Res Vet Med 2008; 6:145150.

    • Search Google Scholar
    • Export Citation
  • 16

    O'Connor T P, Hanscom JL, Hegarty BC, et al. Comparison of an indirect immunofluorescence assay, western blot analysis, and a commercially available ELISA for detection of Ehrlichia canis antibodies in canine sera. Am J Vet Res 2006; 67:206210.

    • Search Google Scholar
    • Export Citation
  • 17

    Landis JR, Koch GG. The measurement of observer agreement for categorical data. Biometrics 1997; 33:159174.

  • 18

    Comer JA, Nicholson WL, Olson JG, et al. Serologic testing for human granulocytic ehrlichiosis at a national referral center. J Clin Microbiol 1999; 37:558564.

    • Search Google Scholar
    • Export Citation
  • 19

    Walls JJ, Aguero-Rosenfeld M, Bakken JS, et al. Inter- and intra-laboratory comparison of Ehrlichia equi and human granulocytic ehrlichiosis (HGE) agent strains for serodiagnosis of HGE by the immunofluorescent-antibody test. J Clin Microbiol 1999; 37:29682973.

    • Search Google Scholar
    • Export Citation
  • 20

    Gomara MJ & Haro I. Synthetic peptides for the immunodiagnosis of human diseases. Curr Med Chem 2007; 14:531546.

  • 21

    Hewer R & Meyer D. Envelope-based HIV vaccine peptides as antigens in HIV-1 immunodiagnostics. Int J Biotechnol 2007; 9:277291.

  • 22

    McCall J W, Genchi C, Kramer LH, et al. Heartworm disease in animals and humans. Adv Parasitol 2008; 66:193285.

  • 23

    Egenvall AE, Hedhammar AA, Bjoersdorff AI. Clinical features and serology of 14 dogs affected by granulocytic ehrlichiosis in Sweden. Vet Rec 1997; 140:222226.

    • Search Google Scholar
    • Export Citation
  • 24

    Egenvall A, Lilliehöök I, Bjöersdorff A, et al. Detection of granulocytic Ehrlichia species DNA by PCR in persistently infected dogs. Vet Rec 2000; 12:186190.

    • Search Google Scholar
    • Export Citation
  • 25

    Beall MJ, Chandrashekar R, Eberts MD, et al. Serological and molecular prevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Ehrlichia species in dogs from Minnesota. Vector Borne Zoonotic Dis 2008; 8:455464.

    • Search Google Scholar
    • Export Citation
  • 26

    Hamer SA, Tsao JI, Walker ED, et al. Use of tick surveys and serosurveys to evaluate pet dogs as a sentinel species for emerging Lyme disease. Am J Vet Res 2009; 70:4956.

    • Search Google Scholar
    • Export Citation
  • 27

    Diniz P P, Beall MJ, Omark K, et al. High prevalence of tick-borne pathogens in dogs from an Indian reservation in northeastern Arizona. Vector Borne Zoonotic Dis 2010; 10:117123.

    • Search Google Scholar
    • Export Citation
  • 28

    Magnarelli LA, Anderson JA. Serologic evidence of canine and equine ehrlichiosis in northeastern United States. J Clin Microbiol 1993; 31:28572860.

    • Search Google Scholar
    • Export Citation
  • 29

    Nyindo MBA, Ristic M & Lewis GE Jr. Immune response of ponies to experimental infection with Ehrlichia equi. Am J Vet Res 1978; 39:1518.

  • 30

    Otranto D, Dantas-Torres F, Breitschwerdt EB. Managing canine vector-borne diseases of zoonotic concern: part one. Trends Parasitol 2009; 25:157163.

    • Search Google Scholar
    • Export Citation
  • 31

    Gaunt SD, Beall MJ, Stillman BA, et al. Experimental infection and co-infection of dogs with Anaplasma platys and Ehrlichia canis: hematologic, serologic and molecular findings. Parasit Vectors 2010; 3:3342.

    • Search Google Scholar
    • Export Citation

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