caa a22 4500 469045000 CHVBK 20180323132821.0 cr unu---uuuuu 170328e19920901xx s 000 0 eng 10.1007/BF01800016 doi (NATIONALLICENCE)springer-10.1007/BF01800016 Mevalonate kinase assay using DEAE-cellulose column chromatography for first-trimester prenatal diagnosis and complementation analysis in mevalonic aciduria [Elektronische Daten] [G. Hoffmann, S. Brendel, S. Scharfschwerdt, Y. Shin, I. Speidel, K. Gibson] Summary: Mevalonic aciduria due to mevalonate kinase deficiency, an inherited defect of cholesterol biosynthesis, has presented with clinical variability in 10 patients from 7 families. We sought to define a genetic basis for this heterogeneity by determining mevalonate kinase activity in fibroblast heterokaryons obtained by polyethylene glycol fusion. To this end we developed a DEAE-cellulose (Cl−) column chromatography procedure for assessing mevalonate kinase in cell extracts that would allow multiple rapid analyses. Fusion of control fibroblasts with those from affected patients from six families with mevalonate kinase deficiency yielded 37% of the mean control activity. None of the fusions between the six cell lines of patients resulted in measurable mevalonate kinase activity. Using the chromatographic procedure, we developed an optimized assay for mevalonate kinase in biopsied chorionic villi.K m values for chorionic villi were similar to those obtained in fibroblasts. Mevalonate kinase activity in biopsied chorionic villi showed a linear increase (0.75-4.3 nmol/min per mg protein) with gestational age from 7 to 14 weeks. Using the optimized assay in biopsied chorionic villi we performed a first-trimester prenatal diagnosis in a pregnancy at risk for mevalonate kinase deficiency and correctly diagnosed an unaffected fetus. The availability of an optimized assay for mevalonate kinase in biopsied chorionic villi should allow reliable first-trimester prenatal diagnosis for families at risk. SSIEM and Kluwer Academic Publishers, 1992 Hoffmann G. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut Brendel S. Department of Neuropediatrics, University of Göttingen, Germany aut Scharfschwerdt S. Department of Neuropediatrics, University of Göttingen, Germany aut Shin Y. Children's Hospital, University of Munich, Germany aut Speidel I. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut Gibson K. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut Journal of Inherited Metabolic Disease Kluwer Academic Publishers 15/5(1992-09-01), 738-746 0141-8955 15:5<738 1992 15 10545 https://doi.org/10.1007/BF01800016 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01800016 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hoffmann G. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut NATIONALLICENCE 700 1- Brendel S. Department of Neuropediatrics, University of Göttingen, Germany aut NATIONALLICENCE 700 1- Scharfschwerdt S. Department of Neuropediatrics, University of Göttingen, Germany aut NATIONALLICENCE 700 1- Shin Y. Children's Hospital, University of Munich, Germany aut NATIONALLICENCE 700 1- Speidel I. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut NATIONALLICENCE 700 1- Gibson K. Department of Neuropediatrics, University of Heidelberg, W-6900, Heidelberg, Germany aut NATIONALLICENCE 773 0- Journal of Inherited Metabolic Disease Kluwer Academic Publishers 15/5(1992-09-01), 738-746 0141-8955 15:5<738 1992 15 10545 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer