Development and evaluation of an automated dye-binding assay for protein in cerebrospinal fluid. 1988

C M Huang
Clinical Pathology Department, Clinical Center, National Institutes of Health, Bethesda, MD 20892.

Addition of sodium dodecyl sulfate (SDS) to Coomassie Brilliant Blue reagent equalizes the binding variability of the dye to various proteins and markedly improves the accuracy of quantification of protein in cerebrospinal fluid. In the presence of SDS, the absorption spectrum and the absorption maximum are affected by reaction time and temperature, age of the dye preparation, and protein constituent. I automated this procedure, to optimize precision, enable use of a smaller sample, decrease hands-on time, maintain consistency in the time of reading, and avoid carryover. The results (y) compared well with those of the aca (x), with a Deming de-biased regression equation of y = 0.991x + 14.1 mg/L, Sy.x = 33.4 mg/L. The within-run and between-run precision (CV) was less than 2.5% and less than 4.5%, respectively. Commonly used antibiotics, flucytosine, or amphotericin B do not interfere. This automated procedure is fast and accurate and requires only 10 microL of sample.

UI MeSH Term Description Entries
D009391 Nephelometry and Turbidimetry Chemical analysis based on the phenomenon whereby light, passing through a medium with dispersed particles of a different refractive index from that of the medium, is attenuated in intensity by scattering. In turbidimetry, the intensity of light transmitted through the medium, the unscattered light, is measured. In nephelometry, the intensity of the scattered light is measured, usually, but not necessarily, at right angles to the incident light beam. Turbidimetry,Nephelometry,Turbidimetry and Nephelometry
D012016 Reference Values The range or frequency distribution of a measurement in a population (of organisms, organs or things) that has not been selected for the presence of disease or abnormality. Normal Range,Normal Values,Reference Ranges,Normal Ranges,Normal Value,Range, Normal,Range, Reference,Ranges, Normal,Ranges, Reference,Reference Range,Reference Value,Value, Normal,Value, Reference,Values, Normal,Values, Reference
D002556 Cerebrospinal Fluid Proteins Proteins in the cerebrospinal fluid, normally albumin and globulin present in the ratio of 8 to 1. Increases in protein levels are of diagnostic value in neurological diseases. (Brain and Bannister's Clinical Neurology, 7th ed, p221) Proteins, Cerebrospinal Fluid,Fluid Proteins, Cerebrospinal
D004396 Coloring Agents Chemicals and substances that impart color including soluble dyes and insoluble pigments. They are used in INKS; PAINTS; and as INDICATORS AND REAGENTS. Coloring Agent,Dye,Dyes,Organic Pigment,Stain,Stains,Tissue Stain,Tissue Stains,Organic Pigments,Pigments, Inorganic,Agent, Coloring,Inorganic Pigments,Pigment, Organic,Pigments, Organic,Stain, Tissue,Stains, Tissue
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001322 Autoanalysis Method of analyzing chemicals using automation. Autoanalyses
D012394 Rosaniline Dyes Compounds that contain the triphenylmethane aniline structure found in rosaniline. Many of them have a characteristic magenta color and are used as COLORING AGENTS. Fuchsins,Magentas,Fuchsin,Triphenylmethane Aniline Compounds,Aniline Compounds, Triphenylmethane,Compounds, Triphenylmethane Aniline,Dyes, Rosaniline
D012967 Sodium Dodecyl Sulfate An anionic surfactant, usually a mixture of sodium alkyl sulfates, mainly the lauryl; lowers surface tension of aqueous solutions; used as fat emulsifier, wetting agent, detergent in cosmetics, pharmaceuticals and toothpastes; also as research tool in protein biochemistry. Sodium Lauryl Sulfate,Irium,Dodecyl Sulfate, Sodium,Lauryl Sulfate, Sodium,Sulfate, Sodium Dodecyl,Sulfate, Sodium Lauryl

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