PubMed · 10222373
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D S Young. 1999. Issues in genetic testing.. https://pubmed.ncbi.nlm.nih.gov/10222373/
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Two rapid and simple procedures for the quantitative analysis of GDP-l-fucose (GDP-Fuc) are described. The methods are based on time-resolved fluorescence and microplate assay technology. The first assay relies on measuring the enzyme activity of alpha1, 3-fucosyltransferase. In this assay, transfer of fucose from GDP-Fuc converts sialyllactosamine to sialyl Lewis x tetrasaccharide, which is detected and quantified by relevant antibodies on a microplate. The formation of the reaction product is directly dependent on the presence of GDP-Fuc in the concentration range of 10-10,000 nM. In the second method GDP-Fuc inhibits the binding of fucose-specific Aleuria aurantia lectin to fucosylated glycan on a microwell. The lectin-based assay is less sensitive than the enzyme assay, but it is cheaper and faster. We used these assays in monitoring the amount of GDP-Fuc in crude lysates of transgenic yeast, which expresses the enzymes producing GDP-Fuc. The newly developed assays are versatile and applicable to measure also other nucleotide sugars or glycosyltransferase activities in a high-throughput manner.
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BACKGROUND: The increasing use of diagnostic tests is probably not only a consequence of increased morbidity. It has been speculated whether some of the increase may be due to inappropriate use of diagnostic tests. MATERIAL AND METHODS: The PubMed database was searched for literature relating to inappropriate use of laboratory tests. Bayes' theorem was used to explain the context dependence of results obtained from diagnostic tests and to calculate predictive values. RESULTS: A theoretical discussion of tests and their use in diagnostic contexts is given, and a method for evaluating the appropriateness of testing is proposed. By calculating predictive values for positive and negative test results, the use of tests can be evaluated in a common probabilistic currency. Inappropriate usage can be defined as testing that does not give probabilistically useful information. INTERPRETATION: The method will be useful for practising physicians as a basis for improving test usage.