Enzyme-linked immunosorbent assays for detection and quantitation of the entomocidal parasporal crystalline protein of Bacillus thuringiensis subspp. kurstaki and israelensis

Abstract

An enzyme-linked immunosorbent assay was used to detect and quantitate the parasporal crystal toxins of Bacillus thuringiensis subspp. kurstaki and israelensis. The assay method described is extremely sensitive, accurate, and highly specific. With this technique, crystalline insecticidal proteins from several subspecies of B. thuringiensis were compared. The dipteran crystal toxin produced by B. thuringiensis subsp. israelensis was shown to share few epitopes with the lepidopteran toxin from B. thuringiensis subspp. kurstaki, tolworthi, berliner, and alesti.

Isolation of upstream control regions of genes expressed at different times during growth and sporulation in Bacillus thuringiensis

“…kurstaki antiserum to subsp. kurstaki crystal protein (Wie et at, 1984). Other authors have reported a similar lack of immunological cross-reactivity between subsp.…”

“…The results of several studies indicated that no significant immunological relationships existed between the mosquitocidal and lepidopteran-toxic crystal proteins. Wie et al (1982) reported that in both Ouchterlony double-diffusion gel analyses and by indirect ELISA, there was no cross-reaction between the two different crystal proteins. However, subsp.…”

In vitro assay for testing the toxicity of Bacillus thuringiensis subsp. israelensis delta endotoxin using blood agarose plates

“…The ,PGel (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) assay (Brussock and Currier 1990) is not reliable for the toxicity assay because the denatured or inactivated toxin shows protein bands on the gel. The enzymelinked immunoassay can be carried out only in highly sophisticated laboratories (Wie et al 1984).…”

Micro-lipid-droplet encapsulation of Bacillus thuringiensis subsp. israelensis delta-endotoxin for control of mosquito larvae

“…Appropriate dilutions were made and assayed for B. thuringiensis subsp. israelensis 5-endotoxin with an enzyme-linked immunosorbent assay (ELISA) procedure (20). Similar procedures to evaluate the efficiency of encapsulation of solubilized toxin yielded erratic results due to inefficient recovery of the proteins from the micro-lipid droplets.…”

“…Sample preparation and ELISA analyses for the B. thuringiensis subsp. israelensis 5-endotoxin were performed as described previously (20) with an antibody raised to the 25to 30-kilodalton crystal proteins.…”