Detection by ELISA of low numbers of Shiga-like toxin-producing in mixed cultures after growth in the presence of mitomycin C Free

Abstract

Summary

Techniques currently available to detect Shiga-like toxin (SLT)-producing lack sensitivity or require specialised equipment and facilities, and in some cases detect only strains belonging to serotype O157. We have used an ELISA technique, capable of detecting both SLTI and SLTII with crude P1 glycoprotein from hydatid cysts, in combination with enhancement of toxin production by culture with mitomycin C. Supernates of Tryptone Soya Broth cultures containing mitomycin C 200 ng/ml were tested for SLTII. For SLTI, cell lysates pre-treated with polymyxin B were tested. In tests with O157:H7 in mixed culture with strain C600 alone, or with C600, and , SLTI could be detected when the proportion of toxigenic organisms represented 1% of the mixture, and SLTII when the proportion was 0‡025%. When faecal samples with added O157:H7 were examined in this system, SLTII-producing strains were detected when they comprised < 0‡1% of the coliform population. This technique is a sensitive and specific assay for detecting low numbers of SLT-producing organisms in mixed culture such as occurs in cases of haemolytic uraemic syndrome and haemorrhagic colitis.

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1992-03-01
2024-03-28
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