[Emergence of linezolid-resistant Enterococcus faecalis strains from two inpatients in a pediatric ward].

We report herein on the isolation of three linezolid-resistant Enterococcus faecalis strains in 2011 from two pediatric inpatients at Kitasato University Hospital, Japan. Three linezolid resistant strains were isolated from two patients who shared the same room of a pediatric inpatient ward. Two linezolid resistant strains were isolated from patient A who had been treated with a total of 17,600mg of linezolid during 60 days of hospitalization (strains 1 and 2). The linezolid resistant E. faecalis persisted through the time that the patient had been discharged from the hospital. Another linezolid resistant strain was isolated from patient B who had no history of linezolid administration. The resistant strain in patient B phased out spontaneously. The minimum inhibitory concentration of linezolid in these strains ranged from 8.0 to 16.0 microg/mL. PCR amplification of the chromosomal gene encoding domain V of the 23S rRNA and subsequent nucleotide sequencing revealed that all the strains had at least one G2576T mutation. The pulse-field-gel electrophoretograms of the DNA treated with the SmaI restriction enzyme showed an identical profile suggesting that they were derived from a single resistant strain. These results suggested that the resistant strain occurred in patient A and was transmitted to patient B within the inpatient ward.

[Annual changes in antimicrobial susceptibility of Campylobacter jejuni and Campylobacter coli isolated from acute diarrheic patients].

Two hundred thirty-one Campylobacter were isolated from acute diarrheic patients between January 2001 and December 2005. We evaluated annual changes in identified species of Campylobacter and their susceptibilities against antibiotics. Campylobacter jejuni (219 strains; 94.8%) and Campylobacter coli (12 strains; 5.2%) were identified to the species. Susceptibilities to four antimicrobial agents, minocycline (MINO), levofloxacin (LVFX), erythromycin (EM) and clindamycin (CLDM) were examined. The resistant rates of four antimicrobial agents in C. coli were significantly higher than that in C. jejuni. The susceptibility of C. jejuni to LVFX was variable, and MICs gave a bimodal distribution. The resistant rate against EM was estimated to be 9.2% in C. jejuni, 66.7% in C. coli. Moreover, young people ranging from 19 to 24 years old were predominant (47.7%) among the Campylobacter enteritis patients.

[Evaluation of antigen diagnostic kit in group A streptococcus mass infection].

We report a Food-borne group A streptococcus epidemic at Kitasato University campus on July 30 and 31, 2005, believed caused by lunch. A current mass group A streptococcus infection differing from the food-borne epidemic above occurred at Kitasato University East Hospital, also believed caused by lunch. Group A streptococcus was detected using a prompt diagnostic kit and bacterial culture from 116 clinical specimens taken from 116 patients with group A streptococcus pharyngitis at Kitasato University East Hospital on August 5, 2005. To investigate the utility of immunochromatographic detection of group A streptococcus antigen, 116 clinical specimens obtained from pharyngeal membranes by swab were examined using a prompt diagnostic kit for group A streptococcus (ImmunoCard STAT! STREP A TEST) and conventional bacterial culture. Group A streptococcus positivity differed between the two methods. Fourteen patients were found to be positive by the prompt diagnostic kit and 23 by bacterial culture. Four patients showing 1.0 x 10(6) cfu/mL estimated by the culture were difficult to diagnose with the prompt diagnostic kit,even though the detection sensitivity of this kit was 1.0 x 10(6) cfu/mL or more. Conventional bacterial culture should therefore be used in addition to the prompt diagnostic kit to detect group A streptococcus, especially in pharyngeal samples obtained from patients with pharyngitis.

Regional variation in the prevalence of extended-spectrum beta-lactamase-producing clinical isolates in the Asia-Pacific region (SENTRY 1998-2002).

We examined the prevalence of extended-spectrum beta-lactamase (ESBL)-producing strains of Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Proteus mirabilis, Citrobacter koseri, and Salmonella spp. that were isolated as part of the SENTRY Asia-Pacific Surveillance Program between 1998 and 2002. During the study period, a total of 6,388 strains were gathered from 17 medical centers in 7 countries and examined for ESBL production and hyperproduction of K. oxytoca chromosomal K1 beta-lactamase enzyme. High rates of confirmed ESBL-producing isolates were found in K. pneumoniae strains from Singapore (35.6%), followed by those from mainland China (30.7%), South Africa (28.1%), and the Philippines (21.9%), whereas the rates were less than 10% in Japan and Australia. ESBL-producing E. coli strains were also prominent in mainland China (24.5%), Hong Kong (14.3%), and Singapore (11.3%). ESBL-producing K. oxytoca were common in the Philippines (38.5%), Singapore (33.3%), and China (30.0%). Hyperproduction of K. oxytoca chromosomal K1 beta-lactamase enzyme was common in Australia and Japan. P. mirabilis strains from Singapore produced ESBL (17.9%) despite the low prevalence (0-8.1%) in other countries. Few ESBL-producing C. koseri and Salmonella spp. strains were found in Japan, Singapore, Taiwan, and South Africa. Although there was variation among countries in substrate preference, ceftazidime was more likely to detect presumptive ESBL phenotype in K. pneumoniae and aztreonam more likely in E. coli, whereas ceftriaxone was the best substrate for the confirmation of ESBL production. ESBL-producing strains showed high levels of co-resistance to aminoglycosides, tetracycline, trimethoprim-sulfamethoxazole, and ciprofloxacin. Imipenem retained activity against all ESBL-producing strains. Organisms expressing ESBLs are widely distributed in the Asia-Pacific region, although prevalence rates vary significantly.

Clinical usefulness of multiplex PCR lateral flow in MRSA detection: a novel, rapid genetic testing method.

Methicillin-resistant Staphylococcus aureus (MRSA) with exogenous cassette DNA containing the methicillin-resistant gene mecA (SCCmec) poses a problem as a drug-resistant bacterium responsible for hospital- and community-acquired infections. The frequency of MRSA detection has recently been increasing rapidly in Japan, and SCCmec has also been classified more diversely into types I-V. A rapid test is essential for early diagnosis and treatment of MRSA infections, but detection by conventional methods requires at least two days. The newly developed multiplex PCR lateral flow method allows specific amplification of femA to detect S. aureus, mecA to detect SCCmec, and kdpC to detect SCCmec type II; moreover, PCR products can be evaluated visually in about 3 h. In the present study, we developed a PCR lateral flow method for MRSA using this method and investigated its clinical usefulness in the detection of MRSA. The results showed a diagnostic concordance rate of 91.7% for MRSA and methicillin-susceptible S. aureus between bacteriological examination and PCR lateral flow, and a high level of specificity in PCR lateral flow. In addition, a higher detection rate for S. aureus using the same sample was observed for PCR lateral flow (70.2%) than for bacteriological tests (48.6%). The above results show that PCR lateral flow for MRSA detection has high sensitivity, specificity, and speed, and its clinical application as a method for early diagnosis of MRSA infections appears to be feasible.