OJVM  Vol.3 No.4 , August 2013
Detection of Antibodies in Serum Specimens from Dogs with Blastomycosis with Lysate Antigens Prepared from Four Blastomyces dermatitidis Dog Isolates: Individual Antigens vs Antigen Combinations
Abstract: Blastomycosis, the systemic fungal infection of humans and animals, has presented a diagnostic challenge to clinicians and laboratory personnel for many years. Our laboratory has been concentrating on attempting to develop antigenic reagents from the yeast phase of various isolates of Blastomyces dermatitidis and to evaluate these lysate antigens with regard to antibody detection in blastomycosis. The aim of this current study was to evaluate yeast phase antigens prepared from four dog isolates of B. dermatitidis and to evaluate their efficacy, when used individually or in combination, for antibody detection in sera from dogs with blastomycosis. Mean absorbance values using the ELISA to assay 24 serum specimens (Trial 1) ranged from 0.588 with an individual lysate antigen to 0.992 when three reagents were combined. Eight of the lysates exhibited mean absorbance values ranging from 0.992 to 0.915 with 7 out of 8 being lysate antigen combinations. Mean absorbance values with the other 6 lysates ranged from 0.899 to 0.588. In Trial 2, the 6 most sensitive reagents from Trial 1 were assayed against 10 highly reactive dog sera. The results of Trial 2 showed that 5 antigen combinations detected antibody to a greater degree than the individual lysate antigen. Combinations of northern and southern antigens were able to detect antibody in serum specimens from either of these geographical regions. Comparative studies are continuing to further evaluate various lysate antigen combinations for antibody detection in blastomycosis.
Cite this paper: J. VanDyke, A. Boyd, J. Sorensen, T. Hine, C. Rayner, A. Zamora and G. Scalarone, "Detection of Antibodies in Serum Specimens from Dogs with Blastomycosis with Lysate Antigens Prepared from Four Blastomyces dermatitidis Dog Isolates: Individual Antigens vs Antigen Combinations," Open Journal of Veterinary Medicine, Vol. 3 No. 4, 2013, pp. 235-239. doi: 10.4236/ojvm.2013.34037.

[1]   M. A. Pfaller and D. J. Diekema, “Epidemiology of Invasive Mycoses in North America,” Critical Reviews in Microbiology, Vol. 36, No.1, 2010, pp. 1-53. doi:10.3109/10408410903241444

[2]   A. F. DiSalvo, “Blastomycosis, in Topley and Wilson’s Microbiology and Microbial Infections,” 9th Edition, Arnold Publishers, London, 1998.

[3]   M. A. DeGroote, R. Bjerke, H. Smith and L. V. Rhodes III, “Expanding Epidemiology of Blastomycosis: Clinical Features and Investigation of 2 Cases in Colorado,” Clinical Infectious Diseases, Vol. 30, No. 3, 2000, pp. 582-584. doi:10.1086/313717

[4]   R. W. Bradsher, “Clinical Features of Blastomycosis,” Seminars in Respiratory Infections, Vol. 12, No. 3, 1997, pp. 229-234.

[5]   R. W. Bradsher, S. W. Chapman and P. G. Pappas, “Blastomycosis,” Infectious Disease Clinics of North America, Vol.17, No. 1, 2003, pp. 21-40. doi:10.1016/S0891-5520(02)00038-7

[6]   J. A. Smith and C. A. Kauffman, “Pulmonary Fungal Infections,” Respirology, Vol. 17, No. 6, 2012, pp. 913-926. doi:10.1111/j.1440-1843.2012.02150.x

[7]   J. R. Bariola and K. S. Vyas, “Pulmonary Blastomycosis,” Seminars in Respiratory Critical Care Medicine, Vol. 32, No. 6, 2011, pp. 745-753. doi:10.1055/s-0031-1295722

[8]   J. A. McKinnell and P. G. Pappas, “Blastomycosis: New Insights into Diagnosis, Prevention, and Treatment,” Clinical Chest Medicine, Vol. 30, No. 2, 2009, pp. 227-239. doi:10.1016/j.ccm.2009.02.003

[9]   M. Saccente and G. L. Woods, “Clinical and Laboratory Update on Blastomycosis,” Clinical Microbiology Reviews, Vol. 23, No. 2, 2010, pp. 367-381. doi:10.1128/CMR.00056-09

[10]   J. E. Cutler, G. S. Deepe Jr. and B. S. Klein, “Advances in Combating Fungal Diseases: Vaccines on the Threshold,” Nature Review of Microbiology, Vol. 5, No. 1, 2007, pp. 13-18. doi:10.1038/nrmicro1537

[11]   K. S. Vyas, J. R. Bariola and R. W. Bradsher, “Advances in the Serodiagnosis of Blastomycosis,” Current Fungal Infection Reports, Vol. 2, No. 4, 2008, pp. 227-231. doi:10.1007/s12281-008-0033-z

[12]   B. S. Klein and J. M. Jones, “Isolation, Purification and Radiolabeling of a Novel Surface Protein on Blastomyces dermatitidis Yeasts to Detect Antibody in Infected Patients,” Journal of Clinical Investigation, Vol. 85, No. 1, 1990, pp. 152-161. doi:10.1172/JCI114406

[13]   P. Connolly, C. A. Hage, J. R. Bariola, E. Bensadoun, M. Rodgers, R. W. Bradsher and J. J. Wheat, “Blastomyces dermatitidis Antigen Detection by Quantitative Enzyme Immunoassay,” Clinical Vaccine Immunology, Vol. 19, No. 1, 2012, pp. 53-56. doi:10.1128/CVI.05248-11

[14]   C. A. Hage, T. E. Davis, L. Egan, M. Parker, D. Fuller, A. M. LeMonte, D. Durkin, P. Connelly, L. J. Wheat, D. Blue-Hindy and K. A. Knox, “Diagnosis of Pulmonary Histoplasmosis and Blastomycosis by Detection of Antigen in Bronchoalveolar Lavage Fluid Using an Improved Second-Generation Enzyme-Linked Immunoassay,” Respiratory Medicine, Vol. 101, 2007, pp. 43-47. doi:10.1016/j.rmed.2006.04.017

[15]   B. S. Klein, R. A. Squires, J. K. Lloyd, D. R. Ruge and A. M. Legendre, “Canine Antibody Response to Blastomyces dermatitidis WI-1 Antigen,” American Review of Veterinary Research, Vol. 61, No. 5, 2000, pp. 554-558. doi:10.2460/ajvr.2000.61.554

[16]   D. Spector, A. M. Legendre, J. Wheat, D. Bemis, B. Rohrbach, J. Taboada and M. Durkin, “Antigen and Antibody Testing for the Diagnosis of Blastomycosis in Dogs,” Journal of Veterinary Internal Medicine, Vol. 22, No. 4, 2008, pp. 839-843. doi:10.1111/j.1939-1676.2008.0107.x

[17]   J. R. Bariola, C. A. Hage, M. Durkin, E. Bensadoun, P. O. Gubbins, L. J. Wheat and R. W. Bradsher, “Detection of Blastomyces dermatitidis Antigen in Patients with Newly Diagnosed Blastomycosis,” Diagnostic Microbiology and Infectious Diseases, Vol. 69, No. 2, 2011, pp. 187-191. doi:10.1016/j.diagmicrobio.2010.09.015

[18]   S. M. Johnson and G. M. Scalarone, “Preparation and ELISA Evaluation of Blastomyces dermatitidis Yeast Phase Lysate Antigens,” Diagostic Microbiology and Infectious Diseases, Vol. 11, No. 2, 1989, pp. 81-86. doi:10.1016/0732-8893(88)90076-4

[19]   J. L. Bono, A. M. Legendre and G. M. Scalarone, “Detection of Antibodies and Delayed Hypersensitivity with Rotofor Preparative IEF Fractions of Blastomyces dermatitidis Yeast Phase Lysate Antigens,” Journal of Medical and Veterinary Mycology, Vol. 33, No. 4, 1995, pp. 209-214. doi:10.1080/02681219580000441

[20]   M. A. Fisher, J. L. Bono, R. O. Abuodeh, A. M. Legendre and G. M. Scalarone, “Sensitivity and Specificity of an Isoelectric Focusing Fraction of Blastomyces dermatitidis Yeast Lysate Antigen for the Detection of Canine Blastomycosis,” Mycoses, Vol. 38, No. 5-6, 1995, pp. 177-182. doi:10.1111/j.1439-0507.1995.tb00046.x

[21]   R. C. Axtell and G. M. Scalarone, “Serological Differences in Three Blastomyces dermatitidis Strains,” Mycoses, Vol. 45, No. 11-12, 2002, pp. 437-422. doi:10.1046/j.1439-0507.2002.00804.x

[22]   J. F. Shurley, A. M. Legendre and G. M. Scalarone, “Blastomyes dermatitidis Antigen Detectioni in Urine Specimens from Dogs with Blastomycosis Using a Competitive Binding Inhibition ELISA,” Mycopathologia, Vol. 160, No. 2, 2005, pp. 137-142. doi:10.1007/s11046-005-3153-9

[23]   J. F. Shurley and G. M. Scalarone, “Isoelectric Focusing and ELISA Evaluation of a Blastomyces dermatitidis Human Isolate,” Mycopathologia, Vol. 164, No. 2, 2007, pp. 73-76. doi:10.1007/s11046-007-9033-8

[24]   C. M. Sestero and G. M. Scalarone, “Detection of IgG and IgM in Sera from Canines with Blastomycosis Using Eight Blastomyces dermatitidis Yeast Phase Lysate Antigens,” Mycopathologia, Vol. 162, No. 1, 2006, pp. 33-37. doi:10.1007/s11046-006-0028-7

[25]   C. M. Sestero and G. M. Scalarone, “Detection of the Surface Antigens BAD-1 and Alpha (1-3) Glucan in Six Different Strains of Blastomyces dermatitidis Using Monoclonal Antibodies,” Journal of Medical and Biological Science, Vol. 1, No. 1, 2006, pp. 1-7.

[26]   W. O. Hatch and G. M. Scalarone, “Comparison of Colorimetric and Chemiluminescent ELISAs for the Detection of Antibodies to Blastomyces dermatitidis,” Journal of Medical and Biological Sciences, Vol. 3, No 1, 2009, pp. 1-6.

[27]   J. C. Wright, T. E. Harrild and G. M. Scalarone, “Sensitivity and Specificity Determinations with Isoelectric Focusing Fractions of Blastomyces dermatitidis for Antibody Detection in Serum Specimens from Infected Dogs,” Open Journal of Vetrinary Meddicine, Vol. 2, No. 4, 2012, pp. 237-241. doi:10.4236/ojvm.2012.24038

[28]   J. J. Roberts, M. V. Madrid, L. Dickerson, B. Hutchison and G. M. Scalarone, “Comparison of Antibody Detection with Yeast Lysate Antigens Prepared from Blastomyces dermatitidis Dog Isolates from Wisconsin and Tennessee,” Open Journal of Veterinary Medicine, Vol. 3, No. 1, 2013, pp. 67-72. doi:10.4236/ojvm.2013.31011

[29]   H. B. Levine, G. M. Scalarone and S. D. Chaparas, “Preparation of Fungal Antigens and Vaccines: Studies on Coccidioides immitis and Histoplasma capsulatum,” Contributions to Microbiology and Immunology, Vol. 3, 1977, pp. 106-125.

[30]   G. M. Scalarone, H. B. Levine and S. D. Chaparas, “Delayed Hypersensitivity Responses of Experimental Animals to Histoplasmin from the Yeast and Mycelial Phases of Histoplasma capsulatum,” Infection and Immunity, Vol. 21, No. 3, 1978, pp. 705-713.