Polymerase chain reaction assay for diagnosis of potentially toxinogenic Corynebacterium diphtheriae strains: correlation with ADP-ribosylation activity assay

Abstract

We have developed a polymerase chain reaction assay for the clinical diagnosis of potentially toxinogenic strains of Corynebacterium diphtheriae, the causative agent of diphtheria. A 910-bp amplification product, overlapping a DNA portion encoding both fragments of the diphtheria toxin, has been found in 28 among the 36 strains tested. In addition, effective toxin production, as evidenced by the ability of bacterial culture supernatants to ADP ribosylate eukaryotic elongation factor 2, was determined. In every case, the presence of an amplification product correlated with an ADP-ribosylation activity, thus confirming the diagnosis. The polymerase chain reaction assay herein described is very rapid (2 h) compared with the Elek immunodiffusion test or the guinea pig lethality test. It can provide a convenient and reliable method for laboratories involved in the identification of toxinogenic corynebacteria.