Actinobacillus actinomycetemcomitans genetic heterogeneity: amplification of JP2-like ltx promoter pattern correlated with specific arbitrarily primed polymerase chain reaction (AP-PCR) genotypes from human but not marmoset Brazilian isolates.
Specific clonal types of Actinobacillus actinomycetemcomitans, a major human periodontal pathogen, may be responsible for clinical manifestations and the production of leukotoxin virulence factors. Leukotoxicity is associated with genetic polymorphism at the promoter region of the leukotoxin (lItx) gene. Here, we describe the use of arbitrarily primed polymerase chain reaction (AP-PCR) and ltx promoter PCR to molecularly characterise 35 A. actinomycetemcomitans Brazilian isolates: 21 of human origin and 14 from captive marmosets (Callitrix spp., primates commonly used as animal models for periodontal research). The discriminative capacity of each of 12 arbitrary primers was found to be variable, yielding between 3 and 24 PCR amplitypes. Combination of the results for all primers led to characterisation of 14 genotypes that grouped into four major clusters based on genetic similarity. Clusters 2, 3, and 4 were discriminative to host origin. A correlation with periodontal disease was suggested for strains belonging to clusters 3 and 4. The JP2-like PCR amplification pattern, associated with highly leukotoxic strains, was exclusive to human isolates and present in 29% of human isolates where it occurred in close relationship with AP genotypes L and J (cluster 3).
Saddi-Ortega
L
L
Departamento de Microbiología, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Carvalho
M A R
MA
Cisalpino
P S
PS
Moreira
E S A
ES
D016976:Aggregatibacter actinomycetemcomitans / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics; D001483:Base Sequence; D001938:Brazil; D002144:Callithrix / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D005098:Exotoxins / Q000235:genetics; D005798:Genes, Bacterial; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014774:Virulence / Q000235:genetics
D001427:Bacterial Toxins; D004269:DNA, Bacterial; D005098:Exotoxins; C018579:leukotoxin