Quantification of Der f 1 in houses of patients allergic to house dust mite, Dermatophagoides farinae, using a locally produced detection reagents.

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dc.contributor.authorSookrung, Nitat
dc.contributor.authorKamlanghan, Thanyarat
dc.contributor.authorIndrawattana, Nitaya
dc.contributor.authorTungtrongchitr, Anchalee
dc.contributor.authorTantilipikorn, Pongsakorn
dc.contributor.authorBunnag, Chaweewan
dc.contributor.authorPattanapanyasat, Kovit
dc.contributor.authorChaicumpa, Wanpen
dc.date.accessioned2012-01-11T10:09:02Z
dc.date.available2012-01-11T10:09:02Z
dc.date.issued2011-03
dc.description.abstractBackground: House dust mite (HDM) allergen quantification in house dust samples before and after the allergen elimination is one means of convincing the target population about the health benefits of allergen removal from their environment. Objective: To produce local reagents for quantification of Der f 1 (major allergen of Dermatophagoides farinae) in dust samples from houses of HDM allergic Thai patients. Methods: Recombinant Der f 1 was used for immunization of a BALB/c mouse for hybridoma production. Polyclonal antibody (PAb) to whole body extract of D. farinae was prepared from an immunized rabbit. A sandwich ELISA (MAb-allergen-PAb) was used, in comparison with the commercialized reagents (Indoor Biotechnology, UK), to quantify Der f 1 in dust samples. Results: Two hybridoma clones, Df1-1 and Df1-2, were established. Their secreted MAbs (MAbDf1-1 and MAbDf1-2, respectively) bound to the homologous antigen as well as native Der f 1 and a crude extract of D. farinae. Epitopes of MAbDf1-1 and MAbDf1-2 were located at amino acid residues 206NSQHYGISNYCQ217 and 283DYW---NSWD-WGDSG298 of Der f 1. MAbDf1-1 had higher affinity to Der f 1 than the MAbDf1-2. A sandwich ELISA (MAbDf1-1-allergen-PAb) and commercialized reagents (MAb1-allergen-MAb2 sandwich ELISA) were used in comparison for quantification of Der f 1 in 42 dust samples collected from bedrooms and living rooms of 21 houses of the HDM allergic patients. All of the 42 dust samples measured by both ELISAs had the Der f 1 levels higher than 2 mg per gram of fine dust which is the HDM allergy sensitizing level. In addition, Der f 1 levels in 41 samples (except 1 sample from a living room) measured by the MAbDf1-1-PAb and MAb1-MAb2 sandwich ELISAs were higher than 10 mg per g of dust which is the morbidity level of HDM allergen. The local sandwich ELISA showed a high coefficient correlation (r = 0.91) in measuring known amounts of recombinant and native Der f 1. The results indicate that the reagents produced in the present study can be used for measuring the environmental levels of HDM Der f 1. The assay can also be used for standardization of the HDM extract for monitoring patient's allergenic status or for immunotherapeutic purpose.en_US
dc.identifier.citationSookrung Nitat, Kamlanghan Thanyarat, Indrawattana Nitaya, Tungtrongchitr Anchalee, Tantilipikorn Pongsakorn, Bunnag Chaweewan, Pattanapanyasat Kovit, Chaicumpa Wanpen. Quantification of Der f 1 in houses of patients allergic to house dust mite, Dermatophagoides farinae, using a locally produced detection reagents. Asian Pacific Journal of Allergy and Immunology, 2011 Mar; 29(1): 78-85en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/136400
dc.language.isoenen_US
dc.source.urihttps://apjai.digitaljournals.org/index.php/apjai/article/viewFile/385/352en_US
dc.subjectHouse dust mite (HDM)en_US
dc.subjectallergenen_US
dc.subjectallergyen_US
dc.subjectDermatophagoides farinaeen_US
dc.subjectDer f 1en_US
dc.subjecthybridomaen_US
dc.subjectmonoclonal antibodyen_US
dc.subjectELISAen_US
dc.subjectrecombinanten_US
dc.subjectquantification test kiten_US
dc.subjectallergic patientsen_US
dc.titleQuantification of Der f 1 in houses of patients allergic to house dust mite, Dermatophagoides farinae, using a locally produced detection reagents.en_US
dc.typeArticleen_US
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