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pre-determined in the same assay in the absence of competing serum) to each well containing serially diluted pooled sera from each group, followed by a 1 h incubation at 37 C

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Pelleted bacterial cells were lysed in 30 ml of lysis buffer (20 mM Tris-Cl (pH 7.5), 0.1% Triton X-100, 1 mg/ml lysozyme) for 20 min and sonicated, and insoluble molecules were removed