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Development of a sensitive solid-phase radioimmunoassay technique for quantification of class-specific Escherichia coli antibodies
Authors:S Stephens
Affiliation:Division of Communicable Diseases, Clinical Research Centre, Watford Road, Harrow, Middlesex, U.K.
Abstract:A specific and sensitive, quantitative solid-phase radioimmunoassay (RIA) has been developed for the detection of Escherichia coli antibodies in serum and secretions. Preparation of affinity-purified anti-E. coli standards allows accurate quantification of G, M and A classes of antibody down to 10 ng/ml using only 20 microliters of sample. This technique has considerable advantages over indirect haemagglutination in sensitivity and accuracy of immunoglobulin class detection. RIA also compares favourably with ELISA in sensitivity and sample size required. Affinity-purified standards may also be used to quantify the ELISA test.
Keywords:Escherichia coli  antibody response  ELISA  quantitative RIA  BSA  bovine serum albumin  ELISA  enzyme-linked immunosorbent assay  IHA  indirect haemagglutination  LPS  lipopolysaccharide  2-ME  2-mercaptoethanol  PBS  phosphate-buffered saline  RIA  radioimmunoassay  RT  room temperature
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