Publication: Cockroach allergen detection and cockroach of allergic Thai patients
Issued Date
2003-03-01
Resource Type
ISSN
0125877X
Other identifier(s)
2-s2.0-0041903683
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Mahidol University
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SCOPUS
Bibliographic Citation
Asian Pacific Journal of Allergy and Immunology. Vol.21, No.1 (2003), 1-9
Suggested Citation
Nitat Sookrung, Pornphan Diraphat, Wanpen Chaicumpa, Pongsri Tongtawe, Yuwaporn Sakolvaree, Pramuan Tapchaisri, Vanna Mahakittikun, Anchalee Tungtrongchitr, Pakit Vichyanond, Chaweewan Bunnag Cockroach allergen detection and cockroach of allergic Thai patients. Asian Pacific Journal of Allergy and Immunology. Vol.21, No.1 (2003), 1-9. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/20921
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Title
Cockroach allergen detection and cockroach of allergic Thai patients
Abstract
Hybridomas secreting monoclonal antibodies (MAb) specific to American cockroach (Periplaneta americana) were produced through a fusion of immune splenocytes of a BALB/c mouse immunized with crude cockroach (CR) extract and mouse myeloma cells. Two hybridomas namely 38G6 and 3C2 were established. These specific hybridomas secreted IgG1 monoclonal immunoglobulins with antigenic specificities to CR protein components of over 207 to 72 kDa and 45 to 40 kDa, respectively. The monoclonal antibodies were applied to select their specific epitopes out of the crude CR extract using affinity chromatography. A Prausnitz-Kustner test revealed that these epitopes were allergens which caused wheals and flares of the skin of a guinea-pig previously sensitized with a pool of serum samples from CR allergic patients. The monoclonal antibodies were also used in a capture ELISA to detect specific IgE in serum samples of allergic Thai patients. It was found that 72% and 76% of the patients had IgE antibodies to the epitopes of MAb 38G6 and MAb 3C2, respectively, indicating that the two epitopes are major CR allergens among the CR allergic Thai patients. An antibody-sandwich ELISA was developed for quantitative detection of CR allergens using the two monoclonal antibodies as a capture reagent and rabbit polyclonal antibodies to crude CR extract as a detection reagent. The assay could detect allergenic epitopes contained in as little as 122 pg of crude cockroach extract, and has high potential for direct measurement of the marker allergens in extracts of environmental samples.