Molecular genotyping of isolates of Actinobacillus actinomycetemcomitans by pulsed-field gel electrophoresis after separate digestion with Sse8387I and XhoI

Abstract

Genomic DNA from 18 Japanese clinical isolates of Actinobacillus actinomycetemcomitans was obtained from six periodontitis patients and analyzed using pulsed‐field gel electrophoresis (PFGE) after separate digestion with Sse8387I and with XhoI. Three isolates from an identical patient were found to share an identical PFGE profile, and isolates from distinct patients were found to have PFGE profiles distinctly different from each other. Consequently, the 18 Japanese clinical isolates were discriminated into six distinct genotypes by means of PFGE. The genomic DNA from the other six reference strains (ATCC33384, ATCC43717, ATCC 43718, JCM2434, JCM2435 and Nig‐1) was discriminated into six genotypes by the same PFGE methodology, and these six genotypes were found to be distinctly different from the six genotypes of the 18 Japanese clinical isolates described above. Serotyping demonstrated three PFGE genotypes in the serotype a strains, four the serotype b strains and three the serotype c strains. The present results clearly suggest that the PFGE procedure after separate digestion with Sse8387I and with XhoI has an excellent discriminatory power amongst strains and has a good genotypability for A. actinomycetemcomitans.

The highly leukotoxic JP2 clone of Aggregatibacter actinomycetemcomitans: evolutionary aspects, epidemiology and etiological role in aggressive periodontitis

“…3), Southern blot restriction fragment length polymorphism (RFLP), and PCR. By use of these methods, considerable genetic diversity of A. actinomycetemcomitans has been demonstrated ( Zambon et al ., 1990; DiRienzo et al ., 1990; Han et al ., 1991; Guthmiller et al ., 1993; Slots et al ., 1993; Poulsen et al ., 1994; van Steenbergen et al, 1994; Asikainen et al ., 1995; Saarela et al ., 1993A, 1995; Haubek et al ., 1995 (I), 1996 (II), 1997A (III); He et al ., 1998; Macheleidt et al ., 1999B; Saddi‐Ortega et al ., 2002; Matsuda et al ., 2002).…”

Resistance of fluorescent-labelled Actinobacillus actinomycetemcomitans strains to phagocytosis and killing by human neutrophils

“…Other groups, however, were unable to detect a correlation between resistance to phagocytic killing and leukotoxic activity (Miyasaki et al ., 1986; Kalmar et al ., 1987; Holm et al ., 1993). Discrepancy in data may be attributed to the considerable genetic heterogeneity among A. actinomycetemcomitans strains isolated from periodontal patients, even within each of the six different serotype groups identified to date (Asikainen et al ., 1995; Paju et al ., 2000; Kaplan et al ., 2002; Matsuda et al ., 2002). In light of the conflicting findings regarding phagocytosis and survival of A. actinomycetemcomitans , we have re‐examined the interaction of this periodontopathogen with human neutrophils.…”

Recurrent infective endocarditis due to Aggregatibacter actinomycetemcomitans: reinfection or relapse?

“…(LRB2000-1 and LRB2000-2) and one isolate from 2005 (LRB2005-1)] and three other non-linked clinical isolates of A. actinomycetemcomitans were compared by PFGE using the restriction enzyme XhoI, which has been shown to have excellent discriminatory power amongst A. actinomycetemcomitans isolates (Matsuda et al, 2002). According to Tenover's criteria (Tenover et al, 1995), the strains isolated in 2000 and 2005 were thus closely related since similar genetic patterns with only one band difference were observed (Fig.…”