Teruko Ohkura

Characterization of Group C and G Streptococcal Strains That Cause Streptococcal Toxic Shock Syndrome

ABSTRACT Twelve strains (the largest number ever reported) of group C and G1 streptococci (GCS and GGS, respectively) that caused streptococcal toxic shock syndrome (STSS) were collected and characterized. Eleven strains were identified as Streptococcus dysgalactiae subsp. equisimilis, and one strain was identified as Streptococcus equi subsp. zooepidemicus. We found that it was the first reported case of STSS caused by S. equi subsp. zooepidemicus. Cluster analysis according to the 16S rRNA gene (rDNA) sequences revealed that the S. dysgalactiae strains belonged to clusters I and II, both of which were closely related. The emm types and the restriction patterns of chromosomal DNA measured by pulsed-field gel electrophoresis were highly variable in these strains except BL2719 and N1434. The 16S rDNA sequences and other characteristics of these two strains were indistinguishable, suggesting the clonal dissemination of this particular S. dysgalactiae strain in Japan. As the involvement of superantigens in the pathogenesis of group A streptococcus-related STSS has been suggested, we tried to detect known streptococcal superantigens in GCS and GGS strains. However, only the spegg gene was detected in seven S. dysgalactiae strains, with none of the other superantigen genes being detected in any of the strains. However, the sagA gene was detected in all of the strains except Tokyo1291. In the present study no apparent factor(s) responsible for the pathogenesis of STSS was identified, although close genetic relationships of GCS and GGS strains involved in this disease were suggested.

Use of a Combination of Brushing Technique and the Loop-Mediated Isothermal Amplification Method as a Novel, Rapid, and Safe System for Detection of Helicobacter pylori

ABSTRACT Gastric mucosal biopsy is widely used in the detection of Helicobacter pylori but is associated with a number of problems, including false-negative results due to sampling error and massive bleeding after biopsy. Given the extended period required to culture H. pylori, detection would be further improved by the use of rapid detection methods such as PCR. Here, we developed a rapid, safe, and convenient method for collecting H. pylori which combines endoscopic brushing with the loop-mediated isothermal amplification (LAMP) method. The specificity and sensitivity of LAMP were examined using nine urease-generating non-H. pylori bacterial species, Escherichia coli, Clostridium perfringens, Campylobacter jejuni, Helicobacter hepaticus, and 51 H. pylori strains. Results showed that H. pylori-specific LAMP primers amplified H. pylori DNA only and that the lowest detection limit of the LAMP reaction was 102 CFU. Brushing and biopsy samples taken from 200 patients with peptic ulcer at Nagoya University Hospital and a regional health care center were subjected to both LAMP and culturing. No adverse effects such as severe bleeding or penetration occurred during the procedure. By LAMP assay, 123 patients were confirmed as H. pylori positive when brushing technique samples were assayed, whereas only 100 were positive when biopsy samples were assayed. Culture assay detected H. pylori in 117 patients when it was combined with the brushing technique and in 96 when it was combined with biopsy. Combination of the endoscopic brushing technique with LAMP is considered a useful and safe system for identifying H. pylori infection.

Recurrent vascular catheter-related bacteremia caused by Delftia acidovorans with different antimicrobial susceptibility profiles

An 11-year-old girl with metastatic neuroblastoma developed recurrent bacteremia during sustained neutropenia after autologous peripheral blood transplantation. All febrile episodes of bacteremia were caused by single Delftia acidovorans strain revealed by ERIC-PCR. This strain became resistant to broad-spectrum penicillins and cephalosporins through antibiotic treatments. Removal of the indwelling vascular catheter resulted in resolution of the infection. So far as we know, this is the first report of vascular catheter-related D. acidovorans bacteremia in Japan.

Nationwide epidemiological study revealed the dissemination of meticillin-resistant Staphylococcus aureus carrying a specific set of virulence-associated genes in Japanese hospitals.

To study comprehensive toxin profiles and the chromosomal diversity of current Japanese hospital-associated meticillin-resistant Staphylococcus aureus (HA-MRSA) strains, we conducted PCR-based identification of 28 toxin genes, and staphylococcal cassette chromosome mec (SCCmec) typing and PFGE analysis of 208 MRSA strains isolated from 100 hospitals throughout Japan. Of the tested HA-MRSA strains, 80.3 % were tst-positive. The most frequent toxin gene profile was characterized by the carriage of 13 genes, tst, sec, seg, sei, sel, sem, sen, seo, lukED, hla, hlb, hld and hlg-2. Ninety of the 208 strains had this profile, which was named pattern A. Among the 118 non-pattern A strains, 100 had similar toxin gene profiles, the concordance rates to pattern A of which were more than 80 %. Consequently, 91.3 % of the examined HA-MRSA strains carried similar toxin profiles, although PFGE patterns showed a wide variation. These strains belonged to SCCmec type II, agr II and coagulase type II. We concluded that, unlike MRSA from many other countries, most of the Japanese HA-MRSA strains belonged to, or were related to, a specific group carrying the set of 13 toxin genes, irrespective of chromosomal diversity. In addition, among the 13 toxin genes, the coexistence rates of tst, sec and sel, and those of seg, sei, sem, sen and seo, were higher than for the other toxin genes. High coexistence rates of tst, sec and sel genes suggested the presence of the pathogenicity island SaPIn1 in these strains.

Analysis of Twin-Arginine Translocation Pathway Homologue in Staphylococcus aureus

Staphylococcus aureus releases a large number of exoproteins, including membrane-active proteins and toxins with superantigenic activity involved in pathogenicity. However, the export pathways of exoproteins in S. aureus have not been reported. We analyzed the function of the staphylococcal twin-arginine translocation (Tat) pathway homologue, the presence of which was recently discovered according to the genome database. The amino-acid sequences of the Tat homologues of S. aureus do not have a high similarity with those of Escherichia coli and other bacteria. Constructed tatC-deficient mutants from distinct parent strains showed the same patterns of exoproteins compared with those of parent strains on two-dementional gel electrophoresis, and the amounts of secreted staphylococcal enterotoxins and toxic shock syndrome toxin-1, of which signal peptides have some features often seen in signal sequences of Tat-dependent proteins, did not change with Western blotting analyses. Therefore, it seems that the Tat pathway does not play a major role in the secretion system of S. aureus, but other export pathways may play an important role in toxin secretion. This is the first experimental report showing the influence of the Tat pathway on the secretion of S. aureus.