Direct detection of Shiga-like toxin-producing Escherichia coli in ground beef using the polymerase chain reaction

Abstract

We recently reported the development and assessment of a technique for the detection of Shiga-like toxin-producing Escherichia coli (SLTEC) using the polymerase chain reaction (PCR) and a digoxigenin-11-dUTP-labelled DNA probe. This technique has now been adapted for the direct identification of SLTEC in ground beef. Ground beef homogenates were diluted 1000-fold to reduce the concentration of components which inhibit the thermostable polymerase. Assessment of four different ground beef samples using the PCR detection technique revealed that fat content was a major inhibitory component. As few as 30 SLTEC ml-1 of a ground beef homogenate were detected using the PCR technique, although it was necessary to enrich six of the samples for positive detection. These findings indicate that the PCR detection technique is suitable for the identification of SLTEC directly from contaminated ground beef without isolation of the bacterium or purification of its DNA.