Comparison of antibody detection with Blastomycesdermatitidis yeast lysate antigens in serum specimens from immunized rabbits and infected dogs

Will Christenson, Rachel Horton, Kayla Campbell, Kelly Meacham, Amber Schroeder, Gene M. Scalarone

.

DOI: 10.4236/oji.2011.13009                

Abstract

This present study was designed to evaluate B. dermatitidis antigens, prepared from two isolates (B5896, 597), when the yeast cells were allowed to lyse in distilled water for one day or seven days. The indirect enzyme-linked immunosorbent assay (ELISA) was used to determine the ability of the lysate reagents to detect antibodies in 30 rabbit and 30 dog serum specimens. Mean absorbance values with B5896 lysate antigen ranged from 1.637 (day 1) to 1.461 (day 7) and absorbance values with 597 antigen ranged from 1.579(day 1) to 1.396 (day 7) with the serum specimens from immunized rabbits. Serum specimens from infected dogs yielded absorbance values ranging from 1.672 (day 1) to 1.763 (day 7) with the B5896 and values ranging from 1.909 (day 1) to 1.224 (day 7) with the 597. Optimal reactivity was obtained with the day 1 lysate using both lysate antigens against the rabbit sera and with the 597 antigen against the dog sera. Slightly greater reactivity was evidenced with the day 7 B5896 antigen when the dog sera was tested. Comparative studies are continuing in order to produce an optimal anti-genic preparation for antibody detection in blastomycosis.

Keywords

ELISA; Blastomycesdermatitidis; Antibody Detection; Lysate Antigens

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	DiSalvo, A.F. (1998) Blastomycosis (Chapter 18). In: Ninth, Ed., Topley and Wilson’s Microbiology and Microbial Infections. Arnold Publishers, London, 337-355.
[2]	Klein, B.S. and Jones, J.M. (1990) Isolation, purification, and radiolabeling of a novel surface protein on Blastomycesdermatitidis yeasts to detect antibody in infected patients. Journal of Clinical Investigation, 85, 152. doi:10.1172/JCI114406
[3]	Bromel, C. and Sykes, J.E. (2005) Epidemiology, diagnosis, and treatment of blastomycosis in dogs and cats. Clinical Techniques in Small Animal Practice, 20, 233-239. doi:10.1053/j.ctsap.2005.07.004
[4]	Bradsher, R.W. (2008) Pulmonary blastomycosis. Seminars in Respiratory and Critical Care Medicine, 29, 174-181. doi:10.1055/s-2008-1063856
[5]	Bradsher, R. W. (1997) Clinical features of blastomycosis. Seminars in Respiratory Infections, 12, 229-234.
[6]	Cutler, J.E., DeepeJr, G.S. and Klein, B.S. (2007) Advances in combating fungal diseases: vaccines on the threshold. Nature Reviews Microbiology, 5, 13-18. doi:10.1038/nrmicro1537
[7]	McKinnell, J.A. and Pappas, P.G. (2009) Blastomycosis: New insights into diagnosis, prevention, and treatment. inics In Chest Medicine, 30, 227-239. doi:10.1016/j.ccm.2009.02.003
[8]	Bakerspigel, A., Kane J. and Schaus, D. (1986) Isolation of Blastomycesdermatitidis from an earthen floor in southwestern Ontario, Canada. Journal of Clinical Microbiology, 24, 890-891.
[9]	Kesselman, E.W., Moore, S. and Embil, J.M. (2005) Using local epidemiology tomake a difficult diagnosis: A case of blastomycosis. Canadian Journal of Emergency Medicine, 793, 171-173.
[10]	Axtell, R.C. andScalarone, G.M. (2002) Serological differences in three Blastomycesdermatitidis isolates. Mycoses, 45, 42-43.
[11]	Axtell R.C. andScalarone, G.M. (2002) Serological differences in two Blastomycesdermatitidis isolates from different geographical regions of North America. Mycopathologia, 15, 141-144. doi:10.1023/A:1014599115064
[12]	Durkin, M., Witt, J., Lemonte, A., Wheat, B. and Connolly, P. (2004) Antigen assay with the potential to aid in the diagnosis of blastomycosis. Journal of Clinical Microbiology, 42, 4873-4875. doi:10.1128/JCM.42.10.4873-4875.2004
[13]	Johnson, S.M. and Scalarone, G.M. (1988) Preparation and ELISA evaluation of Blastomycesdermatitidisyeast phase lysate antigens. Diagnostic Microbiology and Infectious Disease, 11, 81-86. doi:10.1016/0732-8893(88)90076-4
[14]	Bradsher, R.W., Chapman, S.W. and Pappas, P.G. (2003) Blastomycosis. Infectious Disease Clinics of North America, 17, 21-40. doi:10.1016/S0891-5520(02)00038-7
[15]	Bono, J.L., Legendre, A.M.and Scalarone, G.M. (1995) Detection of antibodies and delayed hypersensitivity with Rotofor preparative IEF fractions of Blastomycesdermatitidisyeast phase lysate antigens Journal of Medical and Veterinary Mycology, 33, 209-214. doi:10.1080/02681219580000441
[16]	Klein, B.S., Squires, R.A., Foyer Lloyd, J.K., Ruge, D.R. and Legendre, A.M. (2000) Canine antibody response to BlastomycesdermatitidisWI-1 antigen. American Journal of Veterinary Research, 61, 554-558. doi:10.2460/ajvr.2000.61.554
[17]	Vyas, K.S., Bariola, J.R. and Bradsher, R.W. (2008) Advances in the serodiagnosis of blastomycosis. Current Fungal Infection Reports, 2, 227-231. doi:10.1007/s12281-008-0033-z
[18]	Spector, D., Legendre, A.M., Wheat, J., Bemis, D., Rohrbach, B., Taboada, J. and Durkin, M. (2008) Antigen and antibody testing for the diagnosis of blastomycosis in dogs. Journal of Veterinary Internal Medicine, 22, 839-843. doi:10.1111/j.1939-1676.2008.0107.x
[19]	Levine, H.B., Scalarone, G.M. and Chaparas, S.D. (1977) Preparation of fungal antigens and vaccines: Studies on Coccidioidesimmitis and Histoplasmacapsulatum. Contributions to Microbiology & Immunology, 3, 106-125.
[20]	Levine, H.B., Scalarone, G.M., Campbell, G.D., Graybill, R.C. and Chaparas, S.D. (1979)Histoplasmin-CYL, a yeast phase reagent in skin test studies with humans. American Review of Respiratory Disease, 119, 629-636.
[21]	Scalarone, G.M., Levine, H.B., Chaparas, S.D. and Cobb, J.M. (1973) Properties and assay of spherulins from Coccidioidesimmitis in delayed sensitivity responses of animals. Sabouraudia, 11, 222-234. doi:10.1080/00362177385190461
[22]	Scalarone, G.M., Levine, H.B., Pappagianis, D. and Chaparas, S.D. (1974) Spherulin as a complement-fixing antigen in human coccidioidomycosis. American Review of Respiratory Disease, 110, 324-328.
[23]	Sestero, C.M. and Scalarone, G.M. (2006) Detection of IgG and IgM in sera from canines with blastomycosis using eight Blastomycesdermatitidis yeast phase lysate antigens. Mycopathologia, 162, 33-37. doi:10.1007/s11046-006-0028-7
[24]	Shurley, J.F. and Scalarone, G.M. (2007) Isoelectric focusing and ELISA evaluation of a Blastomycesdermatitidis human isolate. Mycopathologia, 164, 73-76. doi:10.1007/s11046-007-9033-8