Sensitivity and Specificity Determinations with Isoelectric Focusing Fractions of Blastomyces dermatitidis for Antibody Detection in Serum Specimens from Infected Dogs

Joshua C. Wright, Terrick E. Harrild, Gene M. Scalarone

Department of Biological Sciences, Idaho State University, Pocatello, USA.

DOI: 10.4236/ojvm.2012.24038                

Abstract

Blastomycosis and histoplasmosis manifest as lung and systemic fungal infections in mammals caused by Histoplasma capsulatum, and Blastomyces dermatitidis. These infections exhibit cross reactivity of antibodies which makes a correct diagnosis potentially elusive. The purpose of this study was to gain an understanding of which isoelectric focusing fractions (Rotofor^TM) of B. dermatitidis were reactive or cross reactive with serum specimens from dogs infected with B. dermatitidis, H. capsulatum, and Cryptococcus neoformans. Three serum specimens from dogs that were infected with B. dermatitidis, two dogs infected with H. capsulatum, and one dog infected with C. neoformans were assayed against the 20 B. dermatitidis Rotofor^TM fractions. Reactivity was determined using the indirect enzyme linked immunoassay (ELISA). Reactivity with B. dermatitidis was found predominantly in the protein fractions 1 - 6, and cross reactivity with H. capsulatum, and C. neoformans sera was found within the B. dermatitidis protein fractions 15 - 19.

Keywords

Isoelectric Focusing; ELISA; Blastomycosis; Lysate Antigen; Antibody Detection

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	A. F. DiSalvo, “Blastomycosis, in Topley and Wilson’s Microbiology and Microbial Infections,” 9th Edition, Arnold Publishers, London, 1998, pp. 337-355.
[2]	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
[3]	J. R. Bariola and K. S. Vyas, “Pulmonary Blastomycosis,” Seminars in Respiratory Critical Care Medicine, Vo. 32, No. 6, 2011, pp. 745-753. doi:10.1055/s-0031-1295722
[4]	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
[5]	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
[6]	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, 2007, pp. 13-18. doi:10.1038/nrmicro1537
[7]	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
[8]	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
[9]	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.
[10]	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
[11]	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
[12]	J. F. Shurley, A. M. Legendre and G. M. Scalarone, “Blastomyes dermatitidis Antigen Detection 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
[13]	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
[14]	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.
[15]	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
[16]	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. 1, 1995, pp. 177-182. doi:10.1111/j.1439-0507.1995.tb00046.x
[17]	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
[18]	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.
[19]	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.
[20]	H. B. Levine, G. M. Scalarone, G. D. Campbell, R. C. Graybill and S. D. Chaparas, “Histoplasmin-CYL, A Yeast Phase Reagent in Skin Test Studies in Humans,” American Review of Respiratory Diseases, Vol. 119, No. 4, 1979, pp. 629-636.