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
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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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