Estandarización de un método microespectrofotométrico para la determinación de la actividad enzimática de la Fosfomanomutasa 2 en una muestra control de Bogotá Colombia

Abstract

Deficiency in glycosylation PMM2-CDG (CDG Ia) is an autosomal recessive disease, caused by damage of the cytoplasmic enzyme phosphomanomutase 2 (E.C.5.4.2.8), which catalyzes the conversion of mannose-1-phosphate to mannose-6 -phosphate, essential for N-glycosylation. This disease is multisystemic in nature characterized by neurological, digestive, nephrological, hepatic, hematological and cardiac failures. In Colombia there are no clinical laboratories that perform the enzymatic test for diagnosis, nor are there studies that report intervals of enzymatic activity, so the objective of this study was to establish the reference values of the specific activity of PMM2. The microspectrophotometric diagnostic method was standardized with 50 samples of voluntary controls, for this the leukocytes were extracted by the dextran-heparin method from whole blood, sonicated to extract the enzyme of interest and the protein content was quantified by the method of Folin-Lowry. To determine the integrity of the samples, the lysosomal enzyme N-Acetylgalactosamine-6-sulfatosulfatase was used as a control. Statistical analyzes allowed us to verify that there are no significant differences between gender or age when finding a p-value 0.12 and 0.6537 respectively, under a significance level of 0.05. A specific activity value was found in the upper limit higher than reported with respect to other investigations. The reference value of enzymatic activity determined in this study was 6,546-48,023 nmol / h. mg of protein. The activity values obtained in this control group will allow to start the diagnosis of PMM2 deficient patients from neonatal age and thus contribute to the Colombian health system, as well as to give a correct management by the medical staff, as well as the family and thus improve The quality of life of these patients.