Typing of the apx toxin gene of Actinobacillus pleuropneumoniae using PCR

: For the determination of the apx toxin genes in Actinobacillus pleuropneumoniae strains, five PCR reactions were developed which allow the detection of the activator and structural genes and the secretion genes of the toxins ApxI, ApxII and ApxIII. The oligonucleotide primers were chosen in order to amplify characteristic parts of the activator and structural genes apxICA, apxIICA and apxIIICA, and the secretion genes apxIBD and apxIIIBD. The annealing temperature of all five reaction was identical in order to allow the five reactions in a single PCR run. The differences in length between the individual amplified gene fragments allowed all product fragments to be separated in a single electrophoresis. This way, a typical toxin gene pattern could be obtained which is characteristic for the five toxin gene groups of A. pleuropneumoniae including serotypes 1, 5a, 5b, 9 and 11 (group 1), serotypes 2, 4, 6 and 8 (group 2), serotype 3 (group 3), serotypes 7 and 12 (group 4), and serotype 10 (group 5). The identification of the toxin genes, which have significance in virulence, enhance and facilitate differentiation of A. pleuropneumoniae and allow the detection of serotypes with an atypical toxin pattern.