Blood ammonia content determination method and blood ammonia diagnosis kit

A diagnostic kit and blood ammonia technology, applied in the field of medical testing and determination, can solve the problems of undiscovered blood ammonia measurement method, high recovery rate, accuracy and precision, etc., and achieve easy operation, high specificity and high test results precise effect

Inactive Publication Date: 2006-05-10
王尔中
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods require alkalization, which is affected by the formation of endogenous ammonia, which affects its accuracy and precision, and is rarely used at present; the ion exchange method is more accurate than the diffusion method, and the CV is 8%-13%; Selective electrode method is to use NH 3 Diffusion to the surface of the electrode, causing the pH of the electrode to change for measurement, the CV of this method is 3.5% - 4.8%, the recovery rate is high, but all need special instruments, not suitable for the specific reality of our country
[0004] Searching Chinese patents, only found that patent application No. 87105593.7 discloses a quick-frozen cup for blood ammonia measurement, but no ideal method for blood ammonia measurement was found

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment one (single dose)

[0071] The blood ammonia diagnostic kit of the present embodiment comprises:

[0072] Buffer 80mmol / l

[0073] Adenosine triphosphate 2mmol / l

[0074] Phosphoenolpyruvate 2mmol / l

[0075] Reduced coenzyme 0.2mmo / l

[0076] Ammonia kinase 4000U / l

[0077] Pyruvate Kinase 1000U / l

[0078] Lactate dehydrogenase 5000U / l

[0079] Stabilizer 50% (total volume)

[0080] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the measured blood ammonia sample to reagent is 1 / 25, and the reaction direction is negative reaction .

[0081] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0082] ATP + ammonia Ammonia kinase Adenosine diphosphate + phosphoramide

[0083] Adenosine diphosphate + phosphoenolpyruvate pyruvate kinase ATP + Pyruvate

[0084] Pyruvate + reduced coenzyme ...

Embodiment 2

[0087] Embodiment two (two doses)

[0088] The blood ammonia diagnostic reagent of the present embodiment has:

[0089] Reagent I

[0090] Buffer 100mmol / l

[0091] Adenosine triphosphate 5mmol / l

[0092] Phosphoenolpyruvate 5mmol / l

[0093] Reduced coenzyme 0.25mmo / l

[0094] Pyruvate kinase 6000U / l

[0095] Lactate dehydrogenase 12000U / l

[0096] Stabilizer 50% (total volume)

[0097] Reagent II

[0098] Buffer 100mmol / l

[0099] Ammonia kinase 7000U / l

[0100] Stabilizer 50% (total volume)

[0101] When measuring blood ammonia content, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, the secondary wavelength of the test is above 405nm, the volume ratio of the blood ammonia sample to the reagent is 1 / 25, and the reaction direction is negative reaction.

[0102] The specific measurement steps are:

[0103] ATP + ammonia Ammonia kinase Adenosine diphosphate + phosphoramide

[0104] Adenosine diphos...

Embodiment 3

[0108] Embodiment three (three doses)

[0109] The blood ammonia diagnostic reagent of the present embodiment is three doses, has:

[0110] Reagent I

[0111] Buffer 120mmol / l

[0112] Adenosine triphosphate 8mmol / l

[0113] Phosphoenolpyruvate 8mmol / l

[0114] Reduced coenzyme 0.3mmo / l

[0115] Stabilizer 20mmo / l

[0116] Reagent II

[0117] Buffer 120mmol / l

[0118] Pyruvate Kinase 10000U / l

[0119] Lactate dehydrogenase 20000U / l

[0120] Stabilizer 50% (total volume)

[0121] Reagent III

[0122]Buffer 120mmol / l

[0123] Ammonia kinase 10000U / l

[0124] Stabilizer 50% (total volume)

[0125] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the measured blood ammonia sample to reagent is 1 / 25, and the reaction direction is negative reaction .

[0126] The specific measurement steps are:

[0127] ATP + ammonia Ammonia kinase Adenosine diphos...

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PUM

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Abstract

The invention relates to a method for determining the content of blood ammonia, and also the reagent kit for blood ammonia diagnosis. The reagent kit comprises cushioning solution, adenosine triphosphate, phosphoenolpyruvate phosphatase, deacidized type coenzyme, ammonia kinase, pyruvate kinase, lactate dehydrogenase, and stabilizer. By mixing sample and reagent of a predetermiend volumetric ratio, generating coupling reaction between them, subjecting the final reactant to biochemiscal analyser, the main wavelength absorbancy variance ratio (speed) can be detected, and the blood ammonia content can thus be measured. The method of the invention can be used to obtain the needed measurement result purely through biochemical analytic instruments, and advantages of the method include higher sensibility, better accuracy, less susceptibility to contamination of internal or external materials, and easy application.

Description

technical field [0001] The invention relates to a method for measuring blood ammonia content, and at the same time, the invention also relates to a blood ammonia diagnosis kit, which belongs to the technical field of medical examination and measurement. Background technique [0002] Ammonia determination methods include micro-diffusion method, ion exchange method, enzymatic method and ammonia electrode method. The most widely used methods are enzymatic method and blood ammonia analyzer analysis method based on ion selective electrode. [0003] The diffusion method is that after the sample is alkalized, NH is released 3 , Titrate the released ammonia with acid, or use the Nessler reaction to form brown-yellow mercuric ammonium iodide for colorimetry. These methods require alkalization, which is affected by the formation of endogenous ammonia, which affects its accuracy and precision, and is rarely used at present; the ion exchange method is more accurate than the diffusion ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
Inventor 王尔中
Owner 王尔中
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