Enzyme-Linked Immunosorbent Assay for Quantifying Antigens of Dermatophagoides farinae and D. pteronyssinus (Acari: Pyroglyphidae) in House Dust Samples

Abstract

Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody as capture and detector antibodies was developed for quantifying antigens of Dermatophagoides farinae Hughes and D. pteronyssinus (Trouessart) mites contained in house dust samples. This monoclonal antibody was directed against mite antigens that were also reactive to immunoglobulin E antibody in all of 10 serum samples obtained from patients allergic to mites. Histological study using fluorescent antibody revealed that the monoclonal antibody was bound to the major part of the D. farinae mite body section including fecal matter and cuticles. The detection limit of the assay system was 0.17 microgram of soluble antigens of both mite species and the antigen amount corresponding to 0.5 mites per microplate well, whether in live or dead mites. This system did not react to antigens of Tyrophagus putrescentiae (Schrank) and Cheyletus malaccensis Oudemans. Slight inhibition of less than or equal to 21.3% was observed with nonspecific substances contained in house dust, such as wool, cotton, human dander, human hair, soil, and biscuit, but no direct ELISA reactions were obtained with any of these materials. In 49 house dust samples, ELISA was significantly correlated with the conventional microscopic observation method.

Infection-Enhancing and -Neutralizing Activities of Mouse Monoclonal Antibodies against Dengue Type 2 and 4 Viruses Are Controlled by Complement Levels

“…One to 2 months after the second immunization, the mice were boosted with dengue virus antigens of the corresponding type (infected mouse brain homogenate; 1 ϫ 10 7 PFU/mouse), and spleen cells were collected 3 to 4 days after the booster immunization. Hybridoma cells were generated essentially as described by Kohler and Milstein (25) with some modifications (28). Briefly, spleen cells were fused to P3U1 cells using polyethylene glycol-4000 (Merck, Darmstadt, Germany).…”

“…A sandwich ELISA was performed as described previously (28). Briefly, microplates sensitized with goat anti-mouse IgG were incubated with serial 10-fold dilutions of test samples, alkaline phosphatase-conjugated goat anti-mouse IgG, and then p-nitrophenyl phosphate.…”

Development and Evaluation of an Enzyme-Linked Immunosorbent Assay for Quantifying Antibodies to Japanese Encephalitis Virus Nonstructural 1 Protein To Detect Subclinical Infections in Vaccinated Horses

“…Monoclonal antibodies to JEV NS1 antigens were generated, based on a method previously described for generating monoclonal antibodies to different antigens (10,15). Briefly, BALB/c mice were immunized repeatedly with lysates of cells stably producing NS1 antigen (ME4 clone) (14), followed by fusion of the mouse spleen cells with the myeloma cell line P3U1.…”

Distribution of Dermatophagoides mite (Acari: Pyroglyphidae) antigens in homes of allergic patients in Japan

“…In order to facilitate the testing of allergenic mites in dust samples, we employed a simple enzyme-linked immunosorbent assay (ELISA; Konishi and Uehara, 1990) for quantifying an antigen ubiquitously distributed in Dermatophagoides mite bodies; this antigen is a suitable target for immunoassay replacing the conventional counting method. We report here that patient homes had higher antigen levels than control homes in bedclothes, such as comforters and pillows.…”