Design and Optimization of Novel Immuno-dipstick Test for Detection of Teliospores of Kamal Bunt (<i>Tilletia indica</i>) of Wheat

Abstract

An immuno-dipstick test was developed for detection of teliospores of Karnal bunt using crude and DE-52

purified anti-teliospore antibodies. For development of this test, intact teliospores and solubilized teliosporic antigen

were coated on nitrocellulose pad of dipsticks. On observation of the nitrocellulose pad under a stereo zoom

microscope, the intact teliospores were found stuck on the pad after coating but got washed away with first washing.

The positive tests after immunoprocessing of dipsticks indicate that surface antigen may tear off from teliospores and

remains bound to nitrocellulose pad. Standardization and optimization of the test was done following antigen

concentration kinetics and antibody dilution analysis as well as antibody incubation time. The test was also

performed for checking cross-reactivity with other fungal pathogens. DE-52 purified antiserum gave partial

reactivity with spores of only <i>Tilletia barclayana </i>and <i>Puccinia recondita. </i>The sensitivity of test was found to be 5

teliospores and 10 ng for solubilized teliosporic antigen.