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Chapter 107 : Immunological Methods in the Diagnostic Allergy Clinical and Research Laboratory

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Abstract:

This chapter overviews clinically used and research methods for the quantification of total and allergen-specific IgE antibodies and allergen-specific IgG antibodies. The performance of immunoassays for these analytes has continued to improve with the availability of new solid-phase matrices, conjugate labeling technology, standardized reference reagents, and data processing methods. One monoclonal antibody-based noncompetitive solid-phase two-site immunoenzymetric assay (IEMA) is described here as the procedure of choice for research investigations of total serum IgE. Because total serum IgE is a regulated analyte as defined by the amended Clinical Laboratory Improvement Act of 1988, licensed clinical laboratories providing this analytical measurement must demonstrate proficiency in an external proficiency survey such as the Diagnostic Allergy (SE) survey conducted by the College of American Pathologists. A modification of the total serum IEMA described above allows the quantification of the level of "free" or non-omalizumab-bound IgE in serum. A 1992 study highlighted that venom-specific IgG levels of <3.5 μg/ml are associated with an increased risk of allergic reactions upon a resting during the first 4 years of immunotherapy with yellow jacket or mixed vespid venoms. In summary, the diagnostic allergy laboratory uses a number of immunoassay methods to measure IgE and IgG antibodies that aid the clinician in the diagnosis and management of individuals with allergic disease. As more is learned about genetic polymorphisms that predispose individuals to allergic disease, molecular biology methods may also be employed in the diagnostic allergy laboratory of the future.

Citation: Hamilton R. 2006. Immunological Methods in the Diagnostic Allergy Clinical and Research Laboratory, p 955-963. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch107

Key Concept Ranking

Allergic Bronchopulmonary Aspergillosis
0.6101877
Humoral Immune Response
0.4836302
Atopic Dermatitis
0.4184144
Allergic Rhinitis
0.41260305
0.6101877
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References

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1. Barbee, R. A.,, M. Halomen,, M. Lebowitz, and , B. Burrows. 1981. Distribution of IgE in a community population sample: correlations with age, sex and allergen skin test reactivity. J. Allergy Clin. Immunol. 68:106111.
2. Bennich, H. H.,, K. Ishizaka,, S. G. O. Johansson,, D. S. Rowe,, D. R. Stanworth, and , W. D. Terry. 1968. Immunoglobulin E: a new class of human immunoglobulin. Immunology 3:323324.
3. Butler, J. E., and , R. G. Hamilton. 1981. Quantitation of specific antibodies: methods of expression, standards, solid phase considerations and specific applications, p. 173198. In J. E. Butler (ed.), Immunochemistry of Solid Phase Immunoassays. CRC Press, Boca Raton, Fla.
4. Dolen, W. K. 2002. The diagnostic allergy laboratory, p. 883890. In N. R. Rose,, R. G. Hamilton, and , B. Detrick (ed.), Manual of Clinical Laboratory Immunology, 6th ed. ASM Press, Washington, D.C.
5. Golden, D. B. K.,, I. D. Lawrence,, R. G. Hamilton,, A. Kagey Sobotka,, M. D. Valentine, and , L. M. Lichtenstein. 1992. Clinical correlation of the venom-specific IgG antibody level during maintenance venom immunotherapy. J. Allergy Clin. Immunol. 90:386393.
6. Hamilton, R. G. 2005. Science behind the discovery of IgE. J. Allergy Clin. Immunol. 115:648652.
7. Hamilton, R. G. 2005. Assessment of indoor allergen exposure. Curr. Allergy Asthma Rep. 5:394401.
8. Hamilton, R. G., and , N. F. Adkinson, Jr. 1979. Solid phase radioimmunoassay for quantitation of antigen-specific IgG in human serum with I-125 protein A from Staphylococcus aureus. J. Immunol. 122:10731079.
9. Hamilton, R. G., and , N. F. Adkinson, Jr. 1980. Quantitation of antigen-specific IgG in human serum. I. Standardization by a Staphylococcus aureus solid phase radioimmunoassay elu-tion technique. J. Immunol. 124:29662971.
10. Hamilton, R. G., and , N. F. Adkinson, Jr. 1985. Naturally-occurring carbohydrate antibodies: interference in solid phase immunoassays. J. Immunol. Methods 77:95108.
11. Hamilton, R. G., and , N. F. Adkinson, Jr. 1997. Immunological tests for diagnosis and management of human allergic disease: total and allergen specific IgE and allergen specific IgG, p. 881892. In N. R. Rose,, E. C. de Macario,, J. D. Folds,, H. C. Lane, and , R. M. Nakamura (ed.), Manual of Clinical Laboratory Immunology, 5th ed. ASM Press, Washington, D.C.
12. Hamilton, R. G., and , N. F. Adkinson, Jr. 2003. Clinical laboratory assessment of IgE-dependent hypersensitivity. J. Allergy Clin. Immunol. 111:687701. (Review.)
13. Hamilton, R. G., and , N. F. Adkinson, Jr. 2004. In vitro assays for IgE mediated sensitivities. J. Allergy Clin. Immunol. 114:213225. (Review.)
14. Hamilton, R. G.,, G. V. Marcotte, and , S. S. Saini. 2005. Immunological methods for quantifying free and total serum IgE levels in allergy patients receiving Omalizumab (Xolair) therapy. J. Immunol. Methods 303:8191.
15. Hiller, R.,, S. Laffer,, C. Harwanegg, et al. 2002. Microarrayed allergen molecules: diagnostic gatekeepers for allergy treatment. FASEB J. 16:414416.
16. Huss, K.,, N. F. Adkinson,, F. Eggleston,, C. Dawson,, M. Van Natta, and , R. G. Hamilton. 2001. House dust mite and cockroach exposure are strong risk factors for positive allergy skin tests in the Childhood Asthma Management Program. J. Allergy Clin. Immunol. 107:4854.
17. Matsson, P., and , R. G. Hamilton. 1997. Evaluation Methods And Analytical Performance Characteristics of Immunological Assays for Human IgE Antibody of Defined Allergen Specificities: Guideline. National Committee for Clinical Laboratory Standards, document 1/LA20-A. NCCLS, Wayne, Pa.
18. Saini, S. S.,, D. W. MacGlashan,, S. A. Sterbinsky,, A. Togias,, D. C. Adelman,, L. M. Lichtenstein, and , B. S. Bochner. 1999. Down-regulation of human basophil IgE and Fc epsilon RI alpha surface densities and mediator release by anti-IgE infusions is reversible in vitro and in vivo. J. Immunol. 162:5624.
19. Wide, L.,, H. H. Bennich, and , S. G. O. Johannson. 1967. Diagnosis by an in vitro test for allergen antibodies. Lancet ii: 11051107.

Tables

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TABLE 1

Analytes measured in the diagnostic allergy laboratory

Citation: Hamilton R. 2006. Immunological Methods in the Diagnostic Allergy Clinical and Research Laboratory, p 955-963. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch107
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TABLE 2

Clinical conditions in which serological determinations may be useful in differential diagnosis

Citation: Hamilton R. 2006. Immunological Methods in the Diagnostic Allergy Clinical and Research Laboratory, p 955-963. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch107

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