C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/42 (2006.01)
Patent
CA 2407446
The invention relates to a method for determining activity of 5'-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a coloured complex by treating it with ammonium molybdate and a reducing agent, colour intensity is measured by a known method, and from the measured value the activity of 5'-nucle- otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5'-nulceotidase, is calculated with a known calculation and/or with the aid of a calibration curve. According to the invention 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP and 5'-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.
Cette invention concerne un procédé permettant de déterminer l'activité de la 5'-nucléotidase, consistant à incuber un échantillon biologique selon un mode et dans des conditions connus per se avec un pentose monophosphate nucléotidique en tant que substrat, à convertir le phosphate inorganique libéré en un complexe coloré par traitement avec du paramolybdate d'ammonium et un agent de réduction, à mesurer l'intensité de couleur selon un procédé connu, et à partir de cette valeur mesurée, à calculer l'activité de la 5'-nucléotidase ou la quantité de phosphate inorganique libéré par unité de temps, en tant que nombre proportionnel à l'activité de la 5'-nucléotidase, ceci au moyen d'un procédé de calcul connu et/ou avec l'aide d'une courbe d'étalonnage. Selon cette invention, des 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP et 5'-TMP sont utilisés en tant que pentoses monophosphates nucléotidiques, et la mesure est réalisée sur chacun de ces six substrats. L'invention concerne également un ensemble de réactifs permettant de réaliser le procédé susmentionné.
Bertha Andras
Tulok Istvan
Fetherstonhaugh & Co.
Pronuk Biotechnologiai Kft.
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