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Production and evaluation of reagents for detection of Histoplasma capsulatum antigenuria by enzyme immunoassay.
Clin Vaccine Immunol. 2007 Jun; 14(6):700-9.CV

Abstract

The detection of urinary Histoplasma capsulatum polysaccharide antigen (HPA) by enzyme immunoassay (EIA) has proven useful for the presumptive diagnosis of histoplasmosis in AIDS patients. Assay limitations include (i) detection of a largely uncharacterized antigen and (ii) difficulty in reproducibly generating antibodies for use in the EIA. To improve antibody production for use in this test and to better understand the antigen being detected, we compared rabbit antibodies elicited using various immunization schedules, routes, and H. capsulatum-derived antigens. Antibodies were evaluated by EIA for their ability to detect purified H. capsulatum C antigen (C-Ag) and antigenuria. Reported as enzyme immunoassay (EI) units (the A(450) with antigen divided by the A(450) without antigen), results demonstrated that intravenous immunization of rabbits with whole, killed yeast-phase cells (yeast-i.v. regimen) produced antibodies giving the highest EI values in the C-Ag EIA (mean EI units +/- standard deviation, 14.9 +/- 0.6 versus 6.4 +/- 0.4 for rabbits immunized with C-Ag versus 2.4 +/- 0.3 for all other regimens combined). Yeast-i.v. antibodies were highly sensitive for the detection of antigenuria in patients with histoplasmosis, as shown by the following results: 12/12 patients compared to 10/12, 6/12, 3/12, and 3/12, respectively, for antibodies from rabbits immunized with (i) C-Ag; (ii) whole, killed yeast-phase cells administered subcutaneously and intramuscularly; (iii) yeast-phase culture filtrates; and (iv) HPA-positive urine. Rabbits immunized using the yeast-i.v. regimen also gave higher peak antibody titers than rabbits immunized by any other regimen (P < 0.03), and their antibodies were most comparable in reactivity to antibodies produced for use in the standard HPA-EIA test (P < 0.001). Therefore, rabbits immunized using the yeast-i.v. regimen produced the most sensitive antibodies with the highest titers for detection of C-Ag and antigenuria in histoplasmosis patients.

Authors+Show Affiliations

Mycotic Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. mlindsley@cdc.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

17428951

Citation

Lindsley, Mark D., et al. "Production and Evaluation of Reagents for Detection of Histoplasma Capsulatum Antigenuria By Enzyme Immunoassay." Clinical and Vaccine Immunology : CVI, vol. 14, no. 6, 2007, pp. 700-9.
Lindsley MD, Holland HL, Bragg SL, et al. Production and evaluation of reagents for detection of Histoplasma capsulatum antigenuria by enzyme immunoassay. Clin Vaccine Immunol. 2007;14(6):700-9.
Lindsley, M. D., Holland, H. L., Bragg, S. L., Hurst, S. F., Wannemuehler, K. A., & Morrison, C. J. (2007). Production and evaluation of reagents for detection of Histoplasma capsulatum antigenuria by enzyme immunoassay. Clinical and Vaccine Immunology : CVI, 14(6), 700-9.
Lindsley MD, et al. Production and Evaluation of Reagents for Detection of Histoplasma Capsulatum Antigenuria By Enzyme Immunoassay. Clin Vaccine Immunol. 2007;14(6):700-9. PubMed PMID: 17428951.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Production and evaluation of reagents for detection of Histoplasma capsulatum antigenuria by enzyme immunoassay. AU - Lindsley,Mark D, AU - Holland,Heather L, AU - Bragg,Sandra L, AU - Hurst,Steven F, AU - Wannemuehler,Kathleen A, AU - Morrison,Christine J, Y1 - 2007/04/11/ PY - 2007/4/13/pubmed PY - 2007/8/3/medline PY - 2007/4/13/entrez SP - 700 EP - 9 JF - Clinical and vaccine immunology : CVI JO - Clin Vaccine Immunol VL - 14 IS - 6 N2 - The detection of urinary Histoplasma capsulatum polysaccharide antigen (HPA) by enzyme immunoassay (EIA) has proven useful for the presumptive diagnosis of histoplasmosis in AIDS patients. Assay limitations include (i) detection of a largely uncharacterized antigen and (ii) difficulty in reproducibly generating antibodies for use in the EIA. To improve antibody production for use in this test and to better understand the antigen being detected, we compared rabbit antibodies elicited using various immunization schedules, routes, and H. capsulatum-derived antigens. Antibodies were evaluated by EIA for their ability to detect purified H. capsulatum C antigen (C-Ag) and antigenuria. Reported as enzyme immunoassay (EI) units (the A(450) with antigen divided by the A(450) without antigen), results demonstrated that intravenous immunization of rabbits with whole, killed yeast-phase cells (yeast-i.v. regimen) produced antibodies giving the highest EI values in the C-Ag EIA (mean EI units +/- standard deviation, 14.9 +/- 0.6 versus 6.4 +/- 0.4 for rabbits immunized with C-Ag versus 2.4 +/- 0.3 for all other regimens combined). Yeast-i.v. antibodies were highly sensitive for the detection of antigenuria in patients with histoplasmosis, as shown by the following results: 12/12 patients compared to 10/12, 6/12, 3/12, and 3/12, respectively, for antibodies from rabbits immunized with (i) C-Ag; (ii) whole, killed yeast-phase cells administered subcutaneously and intramuscularly; (iii) yeast-phase culture filtrates; and (iv) HPA-positive urine. Rabbits immunized using the yeast-i.v. regimen also gave higher peak antibody titers than rabbits immunized by any other regimen (P < 0.03), and their antibodies were most comparable in reactivity to antibodies produced for use in the standard HPA-EIA test (P < 0.001). Therefore, rabbits immunized using the yeast-i.v. regimen produced the most sensitive antibodies with the highest titers for detection of C-Ag and antigenuria in histoplasmosis patients. SN - 1556-6811 UR - https://www.unboundmedicine.com/medline/citation/17428951/Production_and_evaluation_of_reagents_for_detection_of_Histoplasma_capsulatum_antigenuria_by_enzyme_immunoassay_ L2 - http://cvi.asm.org/cgi/pmidlookup?view=long&amp;pmid=17428951 DB - PRIME DP - Unbound Medicine ER -