Liquid double reagent diagnostic reagent kit for determining content of potassium ions in serum and blood plasma

A kit and dual-reagent technology, applied in the field of liquid dual-reagent diagnostic kits, can solve the problems of troublesome testing, insufficient potassium intake, short stability period, etc. Effect

Active Publication Date: 2010-06-02
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] (6) Others: Aldosterone deficiency or long-term use of anti-aldosterone diuretics and familial hyperkalemic periodic paralysis can increase serum potassium
[0012] (1) Potassium intake is insufficient: long-term low potassium diet, fasting or anorexia, etc.
But there are disadvantages: it needs gas source and fire source, and the safety is poor; it needs repeated calibration and stability when starting up, and it is not convenient for individual detection of emergency specimens; it needs specific deionized water or lithium solution in the measurement, which is troublesome to test and difficult to store reagents; Unstable compressed air can directly affect the stability of the flame photometer; the protein and lipid in the sample can change the

Method used

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  • Liquid double reagent diagnostic reagent kit for determining content of potassium ions in serum and blood plasma
  • Liquid double reagent diagnostic reagent kit for determining content of potassium ions in serum and blood plasma
  • Liquid double reagent diagnostic reagent kit for determining content of potassium ions in serum and blood plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage

[0092] Reagent 1

[0093] Tris buffer 100mM

[0094] Enolpyruvate 5mM

[0095] Adenosine diphosphate 5mM

[0096] Magnesium chloride 5mM

[0097] Kryptofix 221 3.0g / L

[0098] NADH 0.5mM

[0099]Glucose 5mM

[0100] Glucose dehydrogenase 10U / L

[0101] MIT 0.5g / L

[0102] Reagent 2

[0103] Tris buffer 10mM

[0104] Lactate dehydrogenase 50KU / L

[0105] Pyruvate kinase 2KU / L

[0106] MIT 0.5g / L

[0107] PEG 6000 1.0g / L

[0108] Set on the automatic biochemical analyzer (Olympus-400): reaction temperature 37°C, reaction time 1.5 minutes, test main wavelength 340nm, test secondary wavelength 410nm. The ratio of the tested potassium ion sample to the reagent is: sample: reagent 1: reagent 2 = 5 μl: 180 μl: 60 μl, and the reaction is a drop reaction.

[0109] First add the sample and reagent 1, the two are automatically mixed in the automatic biochemical analyzer, det...

Embodiment 2

[0135] Add ammonia removal system and surfactant on the basis of Example 1. Enhanced reagent specificity.

[0136] The specific formula is as follows:

[0137] Reagent 1

[0138] Tris buffer 200mM

[0139] Enolpyruvate 5mM

[0140] Adenosine diphosphate 5mM

[0141] Magnesium chloride 5mM

[0142] Glutamate dehydrogenase 10KU / L

[0143] Alpha-Ketoglutarate 10Mm

[0144] Triton X 405 0.5g / L

[0145] Kryptofix 221 3.0g / L

[0146] NADH 0.5mM

[0147] Glucose 5mM

[0148] Glucose dehydrogenase 10U / L

[0149] MIT 0.5g / L

[0150] Reagent 2

[0151] Tris buffer 10mM

[0152] Lactate dehydrogenase 50KU / L

[0153] Pyruvate kinase 2KU / L

[0154] MIT 0.5g / L

[0155] PEG 6000 1.0g / L

[0156] Glycerin 100g / L

[0157] Anti-interference experiment of the kit

[0158] (1) prepare respectively according to the routine technology of this area and contain ascorbic acid, bilirubin, hemoglobin, the sample to be tested of different concentrations of triglyceride, measure the ant...

Embodiment 3

[0170] The accuracy and the precision of embodiment 3 kits of the present invention

[0171] The accuracy and precision of the kit prepared in Example 2 were tested by conventional detection methods in the art.

[0172] 1. Accuracy

[0173] For the quality control solution I with a target value of 3.98 (3.66-4.30) and the quality control solution II with a target value of 6.04 (5.56-6.52), under the same conditions, use the kit to continuously detect the potassium ion concentration of the same quality control solution for 20 The second time, the average value of the detection results is compared with the target value range to detect the accuracy of the kit. The test results of the accuracy of the kit are shown in Table 10:

[0174] 2. Precision

[0175] Under the same conditions, the potassium ion content of the two quality control solutions was continuously detected 20 times by using the kit, and the coefficient of variation of each kit was compared to detect the precision...

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Abstract

The invention relates to a liquid double reagent diagnostic reagent kit for determining the content of potassium ions in serum and blood plasma. By using the reaction principle that phosphoenolpyruvic acid (PEP) can be catalyzed by K+dependency pyruvate kinase (PK) to be converted into pyruvic acid and taking lactate dehydrogenase (LDH) as indicator enzyme by means of NADH, the reagent kit detects the reduction amount of the NADH at 340 nm and obtains the result that the change of the absorbance every minute (delta/min) is directly proportional to the content of K+, thereby establishing an enzyme coupling method for continuously determining the potassium ions. In addition, enzyme and a primer system required by slowly generating reduction type nicotinoyl coenzyme are added in a system andan enzyme-primer-coenzyme slow reaction system is formed in a reagent, so that the coenzyme can be slowly and circularly supplemented and the reagent can be stabilized when the concentration reaches a certain balance. The liquid double reagent diagnostic reagent kit is convenient and concise to use, carries out rapid detection on a common ultraviolet/visible light analyzer or a semiautomatic/fully-automatic chemical analyzer, needs no special or extra instrument and has low cost.

Description

technical field [0001] The invention relates to a liquid double-reagent diagnostic kit for measuring potassium ion content in serum and plasma by enzymatic method. Background technique [0002] Potassium in the human body is the main cation to maintain the physiological activities of cells, it is necessary to maintain the normal osmotic pressure and acid-base balance of the body, participate in the metabolism of sugar and protein, and ensure the normal function of neuromuscular. Potassium ions mostly exist in the cell, and a small amount exists in the extracellular fluid, and the concentration is relatively constant. Potassium in the human body mainly comes from food. The determination of serum potassium salt is actually the determination of potassium ions in extracellular fluid, but potassium ions in the body are constantly exchanged between cells and body fluids to maintain a dynamic balance. Therefore, the level of serum potassium concentration can also indirectly refle...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/32G01N21/31
Inventor 刘希刘瑶
Owner BEIJING STRONG BIOTECH INC
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