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  • 1
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract. Until now the detection of chymotrypsic or polysaccharidic arcs among precipitating systems indicated a positive diagnosis of Farmer's lung disease (FLD). It was shown that the occurrence of chymotrypsic activity is not absolutely correlated with the presence of clinical disease. The presence of different classes of specific precipitating antibodies was looked for and correlated with clinical symptoms in an attempt to find additional positive criteria for the diagnosis of FLD. The enzyme-linked immunofiltration assay (ELIFA) was adapted to detect the different specific antibody classes. 1400 serum samples were studied from 1350 farmers exposed to mouldy hay. Of 1358 samples examined from asymptomatic subjects tested as part of a systematic screening of the profession. 1356 had either no precipitins or have precipitins detected solely against non-chymotrypsic and non-polysaccharidic antigens. Ten specimens with precipitins joined the study. Only two serum samples from two symptomless individuals showed positive serological criteria for FLD as evidenced by the presence of a chymotrypsic band. The remaining forty-two serum samples from twenty-four patients with clinical signs of extrinsic allergic alveolitis had serological criteria for FLD as either chymotrypsic or polysaccharidic arcs. Only specific IgG in asymptomatic subjects was demonstrated, and when a supplementary class of specific antibody (IgM, IgA or IgE) was detected, there was clinical evidence of FLD in all except one case. Thus ELIFA could prove to be a useful tool for the positive diagnosis and prognosis of patients with FLD. In addition this method allows the recognition of a particular functional antigenic fraction of M. faeni by all classes of specific antibodies between different serum samples.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Enzyme-linked immunofiltration assay (ELIFA) for labeling transferred proteins is an interesting and powerful technique for the rapid specific detection (15 min) of proteins immobilized on nitrocellulose or nylon membranes (0.20 and 0.45 μm). ELIFA does not require fastidious handling of the membranes. Saturation, specific labeling and washing procedures are achieved by filtration, controlled by a monitoring unit which regulates the flow rate and ensures excellent specificity, repetition and reproducibility. The recycling by closed circuit or by repetetive inversion of the flow direction offers the advantage of reducing the volumes of expensive reagents while simultaneously increasing the sensitivity of the technique. The detection limit is at least as low as 1-5 ng using directly or indirectly enzymatically labelled probes. ELIFA may be extended to the identification of glycoproteins using specific ligands such as lectins or to the immunocapture of an antigen using specific antibodies immobilized on an activated membrane. ELIFA complements fast separation, by e.g., isoelectric focusing, polyacrylamide gel electrophoresis, or sodium dodecyl sulfate-polyacrylamide gel electrophoresis and accelerated electrotransfer to membranes with rapid detection reducing the total time for separation transfer and detection to less than 2 h.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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