A sensitive reverse ELISA for the measurement of specific IgE to Der p 2, a major Dermatophagoides pteronyssinus allergen.
BACKGROUND
Epidemiologic studies have shown that the presence of IgE antibodies to house dust mite and other indoor allergens is an important risk factor for asthma.


OBJECTIVE
The aim of this study was to develop a reverse ELISA (rELISA) for measuring specific IgE to Der p 2, a major Dermatophagoides pteronyssinus (Dpt) allergen, as a potential tool for followup of allergen immunotherapy.


METHODS
Recombinant Der p 2 allergen or a monoclonal antibody to Der p 2 was used to coat plates in conventional ELISA (cELISA) and rELISA, respectively. Sera from 48 asthmatic patients with positive skin prick test (SPT+) to D. pteronyssinus extract were analyzed for total IgE and specific IgE to Der p 2, and the results were compared with a group of 41 SPT asthmatic and 30 SPT- control subjects.


RESULTS
The sensitivity of the two assays for Der p 2-specific IgE was 3.9 EU/mL and their specificities were confirmed by inhibition tests, in a dose-dependent manner. There was a significant positive correlation between cELISA and rELISA (r = 0.74; P < 0.0001). However, rELISA was more sensitive than was cELISA, regarding both the positive sera percentage (70.8% vs 52.1%) and the Der p 2-specific IgE levels (28.4 vs 4.5 EU/mL) in SPT+ asthmatic patients.


CONCLUSIONS
rELISA has shown to be a sensitive and alternative method for measuring Der p 2-specific IgE without using radioactive techniques. Detection of specific IgE to major allergens and relevant peptides, and identification of B cell epitopes in allergens will provide valuable information for the design of allergen analogs and peptides for immunotherapy.
Silva
D A
DA
Department of Immunology, Microbiology, and Parasitology, Federal University of Uberlândia, MG, Brazil.
Gervásio
A M
AM
Sopelete
M C
MC
Arruda-Chaves
E
E
Arruda
L K
LK
Chapman
M D
MD
Sung
S S
SS
Taketomi
E A
EA
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D039741:Antigens, Dermatophagoides; D001249:Asthma / Q000097:blood / Q000209:etiology / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008925:Mites / Q000276:immunology*; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E