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Method for detecting blood lactic acid in vitro by using chemiluminescence method

A chemiluminescence, in vitro detection technology, applied in chemiluminescence/bioluminescence, biochemical equipment and methods, and analysis by chemical reaction of materials, etc., can solve the problem that the linear range cannot fully meet the clinical detection, and the sensitivity is not as good as chemiluminescence , inconvenience and other problems, to achieve the effect of being conducive to environmental protection, huge social benefits, and cost reduction

Inactive Publication Date: 2010-02-03
福建省洪诚生物药业有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the detection of light absorption, its resolution is not as sensitive as that of chemiluminescence, and the linear range cannot fully meet the needs of clinical detection. For high-concentration samples, it needs to be diluted and measured, which brings some inconvenience.
[0010] 2. The sensitivity is not as good as chemiluminescence

Method used

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  • Method for detecting blood lactic acid in vitro by using chemiluminescence method
  • Method for detecting blood lactic acid in vitro by using chemiluminescence method
  • Method for detecting blood lactic acid in vitro by using chemiluminescence method

Examples

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Embodiment Construction

[0021] The following examples illustrate the implementation of the present invention.

[0022] Option One:

[0023] Reaction principle: lactic acid + O 2 +H 2 O(LO)→Pyruvate+H 2 o 2

[0024] h 2 o 2 + Luminol (HRP) → Luminous

[0025] 1. Reagents:

[0026] 1.1. Lactic acid standard: single or series concentration;

[0027] 1.2. Substrate solution: containing HRP and LO.

[0028] 2. Determination procedure is carried out according to the table below, and the unit of adding sample is μl.

[0029]

[0030] Option II:

[0031] Reaction principle: L-lactic acid + NAD + (LD)→NADH+pyruvate+H +

[0032] Pyruvate + ADP (PK) → ATP +. . . .

[0033] Glycerol+ATP(GK)→Glycerol-3-phosphate+ADP

[0034] Glycerol-3-phosphate+O 2 (GPO)→Dihydroxyacetone phosphate+H 2 o 2

[0035] h 2 o 2 + Luminol (HRP) → Luminous

[0036] 1. Reagents:

[0037] 1.1. Lactic acid standard: single or series concentration;

[0038] 1.2. Mixed enzyme solution, containing ADP and glycer...

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Abstract

The invention discloses a method for detecting blood lactic acid in vitro by using a chemiluminescence method, which is characterized in that LD is utilized to catalyze L-lactic acid and NAD<+> to generate pyruvate and NADH, the pyruvate and ADP generate ATP under the catalysis of pyruvate kinase (PK), ATP and glycerol generate glycerol-3-phosphoric acid and ADP under the catalysis of GK, the glycerol-3-phosphoric acid is acted by GPO to obtain H2O2, and H2O2 is catalyzed by HPR to enable luminol to emit light; the size of light signals is in positive correlation with the concentration of thepyruvate, i.e. the bigger the concentration of the lactic acid is, the stronger the emitted light signals are; the concentration of the lactic acid can be conjectured by recording the light signals; and the lactic acid with the known concentration is used for detecting the light signals to make a dose-response curve, and the content of the lactic acid of an unknown sample can be calculated throughthe curve. In the invention, a chemiluminescence substance replaces a colored substance to achieve the purposes of sensitivity, stability, wide range and safety. The method can be used for preparinga corresponding commercial kit for quantitatively detecting the lactic acid in body fluids such as whole blood, plasma, cerebrospinal fluid, urine, gastric juice and the like.

Description

technical field [0001] The invention relates to a detection method of a medical reagent. In particular, a method for in vitro detection of blood lactate by chemiluminescence. Background technique [0002] The ultra-trace determination method developed by modern biotechnology has greatly promoted the rapid development of various clinical and scientific research work in various fields of biology and medicine, and has made extraordinary contributions to human scientific undertakings and social progress. [0003] In the 1970s, Arakawa first published the technology of using luminol-peroxidase chemiluminescent reaction for enzyme immunoassay analysis. This technology is due to its high sensitivity (up to 10 -18 mol), fast (measurement results can be obtained within 20 minutes), simple (full automation can be realized), long validity period (up to more than one year), beneficial to environmental protection (no radioactive and teratogenic substances), and are generally welcomed. ...

Claims

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Application Information

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IPC IPC(8): G01N21/76C12Q1/26C12Q1/32C12Q1/48
Inventor 王京
Owner 福建省洪诚生物药业有限公司
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