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Determination of inorganic phosphorus and its diagnostic reagent kit

A diagnostic kit and inorganic phosphorus technology, applied in the field of medical testing and determination, can solve the problems affecting the use and promotion, the difference in the accuracy of sensitivity test results, etc.

Inactive Publication Date: 2006-05-31
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with different reagents and testing routes, the sensitivity of detection and the accuracy of detection results will be very different, which directly affects the use and promotion of this method.

Method used

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  • Determination of inorganic phosphorus and its diagnostic reagent kit
  • Determination of inorganic phosphorus and its diagnostic reagent kit
  • Determination of inorganic phosphorus and its diagnostic reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment one (single agent)

[0051] Prepare the diagnostic kit for inorganic phosphorus in serum according to the following components and dosage:

[0052] Glycine buffer 80mmol / l,

[0053] Pyruvate 3mmol / l,

[0054] Phosphoenolpyruvate 2mmol / l,

[0055] thio-NADH 0.2mmol / l,

[0056] Pyruvate oxidase 6000U / l,

[0057] Phosphoenolpyruvate Carboxylase 4000U / l,

[0058] Malate dehydrogenase 8000U / l,

[0059] Ethylene glycol 50% (accounting for the total volume of the reagent).

[0060] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the volume ratio of the tested sample to the reagent is 1:25, and the reaction direction Negative reaction (decrease in absorbance, the same below).

[0061] Add the serum sample and the prepared single dose, and the two are automatically mixed in the analyzer to detect and record the decrease of the absorbance a...

Embodiment 2

[0062] Embodiment two (two doses)

[0063] Prepare the diagnostic kit for inorganic phosphorus in serum according to the following components and dosage:

[0064] Reagent I——

[0065] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0066] Pyruvate 9mmol / l,

[0067] Phosphoenolpyruvate 5mmol / l,

[0068] NADPH 0.25mmol / l,

[0069] Phosphoenolpyruvate carboxylase 7000U / l,

[0070] Malate dehydrogenase 12000U / l,

[0071] Glycerol 50% (accounting for the total volume of reagent I);

[0072] Reagent II——

[0073] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0074] Pyruvate oxidase 9000U / l,

[0075] Ethylene glycol 50% (accounting for the total volume of reagent II).

[0076] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 30°C, reaction time 15 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the ratio of the total volume of the tested sample to reagent I and reagent II is 1:25, the ratio of the amount of reagent I...

Embodiment 3

[0078] Embodiment three (three doses)

[0079] Prepare the diagnostic kit for inorganic phosphorus in serum according to the following components and dosage:

[0080] Reagent I——

[0081] Triethanolamine buffer 120mmol / l,

[0082] Pyruvate 15mmol / l,

[0083] Phosphoenolpyruvate 8mmol / l,

[0084] thio-NADPH 0.3mmol / l,

[0085] Propylene glycol 20mmol / l;

[0086] Reagent II——

[0087] Triethanolamine buffer 120mmol / l,

[0088] Phosphoenolpyruvate Carboxylase 10000U / l,

[0089] Malate dehydrogenase 16000U / l,

[0090] Propylene glycol 50% (accounting for the total volume of reagent II);

[0091] Reagent III——

[0092] Triethanolamine buffer 120mmol / l,

[0093] Pyruvate oxidase 12000U / l,

[0094] Ethylene glycol 50% (accounting for the total volume of reagent III).

[0095] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the total volume of the tes...

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Abstract

The invention is about the measuring method of Inorganic Phosphates and its diagnosis reagent box. Producing carbon dioxide by reacting pyruvate oxidase with pyruvate under the activation ofInorganic Phosphates in the sample of plasma or serum and so on, and producing oxaloacetic acid by reacting carbon dioxide and phosphoenolpyruvate under the existence of phosphoenolpyruvate carboxylase, and then transferring oxidized coenzyme to reduced coenzyme by reacting oxaloacetic acid and malic acid dehydrogenase. Testing the descending range of dominant wave-length340nm absorbance and finally measuring the content of Inorganic Phosphates. This method has high specificity and would not be contaminated by material of internal and exogenous sources, and the result is precise and accurate. Diving the diagnosis reagent box into double-dose or three-dose can reduces the cross interaction of each element, keeps the stability of the reagent and deposits chronically. Using this method can realize the fast testing in common ultraviolet / visible light analyzer or semiautomatic / automatic analyzer and doesní»t require special or additional apparatus, so the cost is low. Thus, this method can be easily promoted and applied in the whole industry.

Description

technical field [0001] The invention relates to a method for measuring inorganic phosphorus and a diagnostic kit prepared by using the method, belonging to the technical field of medical examination and measurement. Background technique [0002] About 87% of the phosphorus in the human body exists in the bones, and the phosphorus in the blood maintains a dynamic balance relationship with the phosphorus in the bones. On the other hand, there is a certain relationship between the concentration of calcium and phosphorus in the blood, and the product of the two is kept within a certain range. In milligrams, the product of the content of calcium and phosphorus in about 100 ml of blood plasma is 35-40 . In normal people, when blood calcium increases, blood phosphorus decreases, and vice versa. When the above product is greater than 40, calcium and phosphorus are deposited in the bone tissue in the form of bone salt; when it is greater than 70, metastatic calcification may occur;...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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