Mouse monoclonal antibodies specific to allergens of HDM (Der p 1 and Der p 2) and American cockroach (Per a 9 or arginine kinase) and rabbit polyclonal antibodies to crude extracts of HDM and CR were produced. The allergen quantification test kits were based on the principle of two-side ELISA (MAb-allergen-PAb). Dust samples can be collected from houses of allergic patients by using standard protocol. Detection and quantification of the allergens in the dust samples can be done by following the test kits instruction manuals. The detection limit of the test kit is 0.1 μg/g of dust which is lower than allergic sensitize level (>2 μg/g of dust).
The current format of the test kit is a sandwich ELISA which takes about 3 hours for completion. The test kit contains specific monoclonal antibodies as the allergen capture reagents. Rabbit polyclonal IgG is used as the allergen detection reagent. Blocking reagent, washing buffer, ELISA plate, secondary antibody (anti-rabbit IgG)-horseradish peroxidase conjugate, enzyme substrate, and enzyme stopping reagents are provided. The researchers are developing these test kits into a convenient format, namely semi-quantitative immunochromatographic test (ICT).
In Southeast Asia, predominant HDM species are Dermatophagoides pteronyssinus and D. farinae and the predominant CR species is Periplaneta americana (American cockroach). HDM and CR are sources of indoor allergens that prevailed in the environment causing human sensitization to a variety of allergic diseases with increasing incidence worldwide. The allergic manifestation can be as severe as asthma with lethality if not treated promptly and properly. Quantification of the allergens in houses of the allergic patients is an important measure for morbidity intervention (prevent provocation of clinical symptoms). Reduction of the levels of the allergens in the environment prevents naive subjects from being sensitized.
The current format of the test kits is aimed for quantification of the allergens in the dust sample collected from the environment such as from bedroom, living room, and bedding. The test kit format is sandwich ELISA which can detect known amount of the major allergens in the samples. This assay can be conveniently performed in hospital or service laboratories. However, the forthcoming ICT test kit can be self-operated (home use).