Reagent kit for monoamine oxidase MAO single-reagent measurement

A technology of monoamine oxidase and kit, applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., to achieve the effect of simple operation, convenient use, and easy promotion and application

Inactive Publication Date: 2009-08-05
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Therefore, in order to overcome the above-mentioned many problems existing in the field of detecting MAO at present, please provide a stable single-reagent test kit for measuring monoamine oxidase activity in serum and plasma

Method used

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  • Reagent kit for monoamine oxidase MAO single-reagent measurement
  • Reagent kit for monoamine oxidase MAO single-reagent measurement
  • Reagent kit for monoamine oxidase MAO single-reagent measurement

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: the preparation of the single reagent of kit

[0060] The present application exemplifies the preparation of three single-reagent kits, the preparation raw materials, preparation steps and the obtained components of the three single-reagents are:

[0061] 1) Preparation of reagents:

[0062] Required raw materials

[0063] name molecular weight purity Manufacturer Tris-base 121.1 99.9％ Beijing Xinjingke Biotechnology Co., Ltd. NaCl 58.44 Analytical pure Beijing Yili Fine Chemicals Co., Ltd. NaN3 65.01 99％ Merck EDTA-2Na 372.24 BSA 98.0％ Amresco ADP—K 501.3 TritonX—100 647 99.0％ Beijing Xinjingke Biotechnology Co., Ltd. α-ketoglutarate 146.1 99.5％ Beijing Xinjingke Biotechnology Co., Ltd. Benzylamine 107.15 99.5％ across D—glucose 180.16 99.8％ Beijing Xinjingke Biotechnology Co., Ltd. GLDI 237u / mg Toyobo NADH 709.9 roche GDH ...

Embodiment 2

[0106] Embodiment 2: the method and step of detecting MAO in the sample with kit

[0107] 1) The detection parameters are:

[0108] Temperature 37°C

[0109] Wavelength 340nm(main)&405nm(secondary)

[0110] Cuvette light path 1.0cm

[0111] Test Method Rate Method

[0112] Reaction Direction Negative Reaction

[0113] 2) The detection steps are:

[0114] First adjust to zero with water for calibration; then add 0.018ml of sample; then add 0.18ml of reagent, incubate at 37°C for 4 minutes, test for 2 minutes, and calculate the change of the average absorbance per minute (ΔA / min) during the test period. The theoretical K value is -1740.

[0115] 3) Normal reference value of MAO in serum: 0-12U / L

Embodiment 3

[0116] Embodiment 3: the anti-interference ability of detection kit I, II, III

[0117]Simultaneously, the performance of 3 kinds of single kits configured in Example 1, i.e. kits I, II, and III, is detected as follows:

[0118] 1. Anti-interference experiment of the kit

[0119] The samples to be tested containing ascorbic acid, bilirubin, and Hb of different concentrations were configured according to conventional techniques in the art respectively, and kits I, II, and III were used to detect the activity of MAO in the sample in the detection instrument, thereby detecting the effects of the three kits on each The anti-interference ability of a kind of interfering substances (each sample was performed in triplicate, and the average value was taken). Test result (mean value) is as shown in table 1:

[0120] Table 1

[0121]

[0122] The results in Table 1 show that when ascorbic acid ≤ 50mg / dL, bilirubin ≤ 40mg / dL or Hb ≤ 200mg / dL, it does not affect the detection of MAO...

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Abstract

The invention discloses a kit of single reagent used for diagnosing monoamine oxidase (MAO) in serum. MAO hydrolyzed benzylamine, butyl amine, amyl amine, Beta-phenylethyl amine and other amine compounds are utilized to produce oxygen; the oxygen reacts with glutamate dehydrogenase in a coupling way; oxidized coenzyme is formed by the reaction of reduced coenzyme; and the activity of sample MAO formed by quantitative reaction is calculated through measuring the descending speed of the absorbance of light with a wavelength of 340 nm; in addition, enzyme and substrate enzyme which can slowly generate reduced nicotinoyl coenzyme are added in the reaction process, so that an enzyme-substrate enzyme-nicotinoyl coenzyme slow reaction system is formed in single reagent and the nicotinoyl coenzyme can be compensated slowly and circularly and further the single reagent can be stabilized when the concentration of the single reagent achieves a certain balance. The kit is convenient for use and simple is structure, can be used on a common ultraviolet / visible light analyzer or a semi-automatic / full-automatic biochemical analyzer for rapid measurement without using a special or additional apparatus and has low cost.

Description

technical field [0001] The application relates to a method for measuring monoamine oxidase activity with an enzymatic single reagent and a preparation method of a kit. Background technique [0002] Monoamine oxidase (MAO) can reflect the biochemical process of fibrosis, and is an important indicator reflecting liver fibrosis and liver cell damage. It has been paid more and more attention in the diagnosis of liver cirrhosis. [0003] MAO is a copper-containing protein that widely exists in the connective tissue of liver, kidney, brain and other organs and in the mitochondria of cells. Its distribution is liver>heart>kidney>brain>lung>skeletal muscle. MAO is also contained in platelets and placenta. There are three isozymes, namely MAO-I, MAO-II and MAO-III, with two subunits MAO-A and MAO-B. Through homologous sequence analysis, it was found that monoamine oxidase has four highly conserved regions. In monoamine oxidase B, these conserved regions include: 1) A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31C12Q1/26
Inventor 温云飞
Owner BEIJING STRONG BIOTECH INC
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