ABSTRACT Dermatophagoides farinae and D. pteronyssinus are the prevalent house dust mites (HDM) in tropical countries and are associated with allergic diseases. This investigation developed a time- resolved immunofluorometric assay (TR-IFMA) for the first time to detect specific IgE antibody in patients with skin prick test positive to HDM but no detectable IgE by other means. Levels of IgE to natural and recombinant HDM allergens were measured by TR-IFMA in 50 HDM-allergic patients and 19 healthy participants compared to sandwich enzyme-linked immunosorbent assay (ELISA). A recombinant allergen, rDerf2, showed a 14 kDa band corresponding to broad range proteins of natural HDM.TR-IFMA showed sensitivity lower than 0.35 kUA/l. TR-IFMA employing three HDM antigens showed good correlations with sandwich ELISA at R2 0.93-0.96. TR-IFMA detected HDM IgE in 62, 62, 25 percent of allergic patient serum sample compared to 28, 32, and 22 percent detected by ELISA result using three HDM allergen. TR-IFMA also detected 26.3, 31.6, and 5.3 percent positive samples from 19 healthy participants while ELISA showed 0, 5.3, and 0 percents IgE positive samples. The use of rDerf2 as an HDM allergen for the assay was verified with no statistically different from other HDM allergens. TR-IFMA showed lower detection limit than ELISA and yielded higher sensitivity for serum of people with allergic symptoms with no detectable HDM IgE. It is anticipated that TR-IFMA for HDM-specific IgE detection will play an important role in future diagnosis of HDM allergy in clinical laboratories and for different research purposes.
Cite this paper
nullPhiphatchaipaisarn, R. , Rabablert, J. , Bramarapravati, K. , Thongdee, D. , Wongpitoon, N. , Durongpisitkul, W. and Malainual, N. (2010) Development of time-resolved immunofluorometric assays for the detection of house dust mite-allergic IgE in human sera. Health, 2, 1280-1286. doi: 10.4236/health.2010.211190.
 Arruda, L.K., Rizzo, M.C., Chapman, M.D., Fernandez- Caldas, E., Baggio, D., Platts-Mills, T.A. and Naspitz, C.K. (1991) Exposure and sensitization to dust mite allergens among asthmatic children in S?o Paulo, Brazil. Clinical & Experimental Allergy, 21(4), 433-439.
Colloff, M.J. (1998) Distribution and abundance of dust mites within homes. Allergy, 53, 24-27.
Nadchatram, M. (2005) House dust mites, our intimate associates. Tropical Biomedicine, 22(1), 23-37.
Thomas, W.R., Smith, W.A. and Hales, B.J. (2004) The allergenic specificities of the house dust mite. Chang Gung Medical Journal, 27(8), 563-569.
Williams, P., Sewell, W.A., Bunn, C., Pumphrey, R., Read, G. and Jolles, S. (2008) Clinical immunology review series: an approach to the use of the immunology laboratory in the diagnosis of clinical allergy. Clinical & Experimental Immunology, 153(1), 10-18.
Bush, R.K., Wood, R.A. and Eggleston, P.A. (1998) La- boratory animal allergy. Journal of Allergy and Clinical Immunology, 102(1), 99-112.
Chapman, M.D., Heymann, P.W., Wilkins, S.R., Brown, M.J. and Platts-Mills, T.A. (1987) Monoclonal immunoassays for major dust mite (Dermatophagoides) allergens, Der p I and Der f I, and quantitative analysis of the allergen content of mite and house dust extracts. Journal of Allergy and Clinical Immunology, 80(2), 184-194.
Ferrandiz, R., Casas, R. and Dreborg, S. (1997) Purification and IgE binding capacity of Der s 3, a major allergen from Dermatophagoides siboney. Clinical & Experimental Allergy, 27(6), 700-704.
Suzuki, M., Tanaka, Y., Korematsu, S., Mikami, B. and Minato, N. (2006) Crystal structure and some properties of a major house dust mite allergen, Derf 2. Biochemical and Biophysical Research Communications, 339(2), 679- 686.
Sheng, S.L., Bao, S.H., Huang, G. and Wang, L.M. (2008) Development of time-resolved immunofluorometric assays for vascular endothelial growth factor and application on plasma of patients with gastric tumours. Clinical & Experimental Immunology, 151(3), 459-466.
Piboonpocanun, S., Malainual, N., Jirapongsananuruk, O., Vichyanond, P. and Thomas, W.R. (2006) Genetic polymorphisms of major house dust mite allergens. Clinical & Experimental Allergy, 36(4), 510-516.
Tanyaratsrisakul, S., Malainual, N., Jirapongsananuruk, O., Smith, W.A., Thomas, W.R. and Piboonpocanun, S. (2009) Structural and IgE binding analyses of recombinant der p 2 expressed from the hosts escherichia coli and pichia pastoris. International Archives of Allergy and Immunology, 29(3), 190-198.
Hales, B.J., Hazell, L.A., Smith, W. and Thomas, W.R. (2002) Genetic variation of Der p 2 allergens: effects on T cell responses and immunoglobulin E binding. Clinical & Experimental Allergy, 32(10), 1461-1467.
Koyanagi, S., Maeda, T., Murakami, T., Kawatsu, K., Sugawara, K., Miyatsu, Y. and Mizokami, H. (2008) Large-scale production of major house dust mite allergen der f 2 mutant (C8/119S) in escherichia coli. Journal of Bioscience and Bioengineering, 106(4), 387-392.
Pittner, G., Vrtala, S., Thomas, W.R., Weghofer, M., Kundi M., Horak, F., Kraft, D. and Valenta, R. (2004) Component-resolved diagnosis of house-dust mite allergy with purified natural and recombinant mite allergens. Clinical & Experimental Allergy, 34(4), 597-603.
Yasueda, H., Mita, H., Yui, Y. and Shida, T. (1989) Com- parative analysis of physicochemical and immunoche- mical properties of the two major allergens from Dermatophagoides pteronyssinus and the corresponding allergens from Dermatophagoides farinae. International Archives of Allergy and Applied Immunology, 88(4), 402- 407.
Parra, M.D., Tuomola, M., Cabezas-Herrera, J. and Ceron, J.J. (2006) Analytical and clinical validation of a time- resolved immunofluorometric assay (TR-IFMA) for canine C-reactive protein in serum. Veterinary Research Communications, 30(2), 113-126.
Yuan, A.S. and Gilbert, J.D. (1996) Time-resolved fluo- roimmunoassay for the determination of lisinopril and enalaprilat in human serum. Journal of Pharmaceutical and Biomedical Analysis, 14(7), 773-781.
Ferrandiz, R., Casas, R., Dreborg, S., Einarsson, R. and Fernandez, B. (1995) Crossreactivity between Dermatophagoides siboney and other house dust mite allergens in sensitized asthmatic patients. Clinical & Experimental Allergy, 25(10), 929-934.
Wu, F.B., Ouyan, H.Q., Tang, X.Y. and Zhou, Z.X. (2008) “Double-antigen sandwich time-resolved immunofluoro- metric assay for the detection of anti-hepatitis C virus total antibodies with improved specificity and sensitivity,” Journal of Medical Microbiology, 57, 947-953.
Sheng, S.L., Wang, Q. and Huang, G. (2007) Development of time-resolved immunofluorometric assays for CA 72-4 and application in sera of patients with gastric tumors. Clinica Chimica Acta, 380(1-2), 106-111.