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Reagent for detecting argininosuccinic acid lyase in serum and preparation method thereof

An arginyl succinate and lyase technology, which is used in the field of reagents for detecting arginyl succinate lyase in serum and the preparation thereof, and can solve the problems of inability to accurately detect ASAL, poor specificity of chronic liver diseases, and the like

Inactive Publication Date: 2018-07-24
湖北海卓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] To sum up, the problems existing in the prior art are: due to the defects of the detection markers themselves, ALT and AST have poor specificity for the detection of chronic liver diseases, and although ASAL has high specificity, due to the methodological The problem, ASAL has been unable to accurately detect

Method used

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  • Reagent for detecting argininosuccinic acid lyase in serum and preparation method thereof
  • Reagent for detecting argininosuccinic acid lyase in serum and preparation method thereof
  • Reagent for detecting argininosuccinic acid lyase in serum and preparation method thereof

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Effect test

preparation example Construction

[0035] like figure 1 As shown, the preparation method of the reagent for detecting arginylsuccinate lyase in serum provided by the embodiment of the present invention comprises the following steps:

[0036] S101: Add 10ul of sample, add R1, 200ul, mix well, and incubate at 37°C for 2-3min;

[0037] S102: Add R2, 50ul, mix well, incubate at 37°C for 1min, continuously measure the absorbance within 3min, and calculate the change rate of absorbance.

Embodiment 1

[0040] The kit for detecting arginylsuccinate lyase in serum according to the embodiment of the present invention includes R1 and R2 reagents, and both R1 and R2 use water as a solvent.

[0041] R1 reagent contains the following components:

[0042]

[0043] R2 reagent contains the following components:

[0044]

[0045] 1. Repeatability determination:

[0046] The same sample was continuously measured 20 times, and the measured average value, standard deviation, and coefficient of variation were as follows:

[0047]

[0048]

[0049] Biochemical detection requires that the coefficient of variation is not greater than 10%, and the coefficient of variation of the present invention is 1.5%, which is far lower than the standard of 10%, so the repeatability of the scheme is good.

[0050] 2. Accuracy detection

[0051] target value

[0052] Biochemical detection requires that the relative deviation of readiness is not more than 10%, and the relative deviat...

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Abstract

The invention belongs to the technical field of detection of argininosuccinic acid lyase, and discloses a reagent for detecting argininosuccinic acid lyase in serum and a preparation method thereof. The reagent consists of R1 and R2, wherein R1 consists of a buffer solution, nicotinamide adenine dinucleotide, fumarase, malic dehydrogenase, an enzyme protective agent, a preservative, a surfactant and ethylenediaminetetraacetic acid, and R2 consists of a buffer solution, disodium arginine succinate, ethylenediaminetetraacetic acid and hydrazine sulfate. The method comprises: adding 10ul of a sample, adding 200ul of R1, mixing the sample and R1 uniformly, and carrying out incubation at 37 DEG C for 2-3min; and adding 50ul of R2, mixing the components uniformly, carrying out incubation at 37 DEG C for 1 min, then continuously measuring the absorbance in 3min, and calculating an absorbance change rate. According to the invention, fumarase, another product generated by catalyzing argininosuccinic acid by ASAL, is detected, and the amount of ASAL is detected by a full-automatic biochemical instrument by utilizing the characteristic that NAD+ has an absorption peak at a 340nm position.

Description

technical field [0001] The invention belongs to the technical field of arginylsuccinate detection, and in particular relates to a reagent for detecting arginylsuccinate lyase in serum and a preparation method thereof. Background technique [0002] Arginylsuccinate (ASAL), also known as arginylsuccinate lyase, is one of the important enzymes in the hepatic urea cycle, which catalyzes the production of arginine and fumarate from argininosuccinate. ASAL is mainly expressed in human liver, kidney and red blood cells. Although ASAL was expressed in the kidney, serum ASAL activity was not increased in renal patients, but significantly increased in hepatitis patients. The two most important indicators for the traditional detection of chronic liver disease are alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The sensitivity of (ALT) to diagnose chronic liver disease is 97.6%, and the specificity is 24.7%; the sensitivity of aspartate aminotransferase (AST) to d...

Claims

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Application Information

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IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 温云飞王成林黄冬琴
Owner 湖北海卓生物科技有限公司
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