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Acetylcholinesterase analysis

A technology of acetylcholinesterase and acetylcholine iodide, which is applied in the field of saliva-based analysis, and can solve problems such as endangering public health, residual chemicals and the like

Inactive Publication Date: 2004-06-30
INST FOR MEDICAL RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Indiscriminate use of pesticides to protect crops from pests and diseases before crops are harvested has the potential to leave large amounts of chemicals and endanger public health

Method used

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  • Acetylcholinesterase analysis

Examples

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Effect test

Embodiment 1

[0016] Example 1 Macroscopic Analysis of Enzyme Activity

[0017] Macroscopic analysis was performed on 12 subjects, 6 males and 6 females. The subjects were selected according to the method of Moss et al. (1994). All 12 subjects were healthy adults, and none of them were at risk of changing cholinesterase levels: for example, long-term exposure to pesticides, acute infections and effects on the heart and lungs , chronic diseases of the kidney, brain or endocrine system, genetic diseases, symptoms of mental illness or recent surgical treatment.

[0018] Saliva samples were obtained from 12 subjects, and filter paper was used to remove air bubbles and debris such as sputum. The 12 cases of saliva samples were stored in an ice box. Transfer the sample to a plastic vial with a plastic tip and centrifuge at 8,000 rpm for 10 minutes at 4°C. In the meantime, prepare a potassium phosphate buffer at pH 6.8. The composition of the buffer is: 0.4735g Na 2 HPO 4 Solution in 50ml ...

Embodiment 2

[0020] Example 2 Microscopic Analysis of Enzyme Activity

[0021] The above experiments were repeated with microscopic analysis samples. The experimental method is the same as the method for macroscopic analysis of acetylcholinesterase, but the dosage is different. The respective volumes of potassium buffer, substrate and DTNB were not 333 μl but 50 μl. Saliva samples were obtained from the original 12 subjects. First, 50 µl of phosphate buffer was added to 50 µl of substrate and 50 µl of DTNB, and placed in one well of a microtiter plate, thereby preparing a blank. The optical density of the blank control was 0.08. 50 μl of substrate and 50 μl of DTNB were pipetted into the other 12 wells of the above microtiter plate. Then, 50 μl of centrifuged saliva was added to each well, and incubated at room temperature (25° C.) for 30 minutes. After 30 minutes, the optical density was read with an immunoassay reader at a wavelength of 410 nm. The results are shown in Table 3. ...

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Abstract

A method for the assay of acetylcholinesterase in a sample of saliva is provided. The method can be used for the detection of poisoning caused by carbamate insecticide or organic phosphorus-based agricultural chemicals in a patient. A kit using the method of the invention is also provided.

Description

technical field [0001] The present invention relates to a saliva-based assay for the detection of human acetylcholinesterase (AChE). Background technique [0002] Cholinesterase (ChE) is a collective term for a group of related enzymes that hydrolyze choline esters. There are two kinds of cholinesterase in the body, and their enzymatic properties, physiological functions and distribution in the body are different. The first is acetylcholinesterase (also known as true cholinesterase), which specifically degrades acetylcholine and is abundantly present in erythrocytes (erythrocytes), nervous tissue, muscle, etc., and its distribution is associated with these physiological functions . Another cholinesterase is butylcholinesterase (also known as pseudocholinesterase), which acts on cholines, such as benzoylcholine and butylcholine, which are found in large amounts in serum and liver , produced in the liver, its physiological effects are thought to be most likely related to th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/46
CPCC12Q1/46
Inventor 李汉林纳兹尼·W·阿迈德阿普杜拉·A·加尼
Owner INST FOR MEDICAL RES
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