Rapid detection of Shigella dysenteriae and Shigella flexneri in faeces by an immunomagnetic assay with monoclonal antibodies

Abstract

A rapid and sensitive method for the detection of Shigella dysenteriae type 1 and Shigella flexneri serotypes in faeces based on capture of the bacteria with immunomagnetic particles is described. The particles were coated with either of two different monoclonal antibodies specific for the O-antigens of Shigella dysenteriae type 1 and Shigella flexneri serotypes. Captured bacteria were detected by an enzyme immunoassay with O-antigen specific rabbit antiserum. The whole assay required 2 to 3 hours to perform and the sensitivity limit was 10(3) cfu/ml as determined by viable cell counting. One hundred and fifty enterobacteria strains, including 100 Shigella strains from a strain collection, and 302 fresh faecal samples were used for the study. All Shigella dysenteriae type 1 and Shigella flexneri culture-positive faecal samples were positive in the immunomagnetic assay. In addition 18 of 252 culture-negative faecal samples were positive. The immunomagnetic assay was compared with latex agglutination and indirect immunofluorescence using culture as the reference method. The immunomagnetic assay had a sensitivity of 100%, latex agglutination a sensitivity of 72% with 28% false-negative results, and indirect immunofluorescence a sensitivity of 95%. The immunomagnetic assay was superior in sensitivity since it also detected Shigella in faecal samples up to two days after antibiotic therapy had been started, which both latex agglutination and indirect immunofluorescence failed to do. The high sensitivity in detecting live and dead bacteria, and the ease of performance of the immunomagnetic assay render it an attractive method for detection of Shigella