Institute for Medical Research

Wasi Ahmad Nazni

Han Lim Lee

A method for checking for sensitivity of acetylcholine esterase in insects using a single insect, includes homogenizing a single insect in aqueous, neutral to acid pH phosphate buffer, providing a solution of acetylcholine iodide in neutral to acid pH phosphate buffered water-miscible organic solvent, providing a solution of 5,5-dithio-bis-(2-nitrobenzoic acid) in the buffer, and providing a solution of propoxur in the buffer, dropping the insect homogenisate into the wells of an assay plate, dropping the acetylcholine iodide and 5,5-dithio-bis-(2-nitrobenzoic acid) solutions into some of the wells and 5,5-dithio-bis-(2-nitrobenzoic acid) propoxur solutions into the other wells and checking for a difference in the yellow coloration between the samples in the two sets of cells.

Method for detecting sensitivity to acetylcholine esterase in insects

A method for checking for elevated activity of mixed-function oxidases using a single insect, includes homogenizing a single insect in an acid pH buffer, preferably by grinding it in a small tube, such as an Eppendorf tube, with a pestle, adding a buffered tetramethylbenzidine solution as substrate, adding hydrogen peroxide as oxidant, and checking the blue coloration, the intensity of which indicates the degree of elevated mixed function oxidase activity. The invention also provides a test kit for use in the method which comprises separate containers of: I. an acid pH buffer, preferably an acid buffer about pH5 II. tetramethylbenzidine III. a solvent for tetramethylbenzidine, preferably methanol, and IV. hydrogen peroxide.

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Selangor, Malaysia

Wasi Ahmad Nazni