Kyoung Ho Roh

Investigation of Toxin Gene Diversity, Molecular Epidemiology, and Antimicrobial Resistance of Clostridium difficile Isolated from 12 Hospitals in South Korea

BACKGROUND Clostridium difficile is a major cause of antibiotic-associated diarrhea. The objective of this study was to characterize clinical isolates of C. difficile obtained from various regions in Korea with regard to their toxin status, molecular type, and antimicrobial susceptibility. METHODS We analyzed a total of 408 C. difficile isolates obtained between 2006 and 2008 from 408 patients with diarrhea in 12 South Korean teaching hospitals. C. difficile toxin genes tcdA, tcdB, cdtA, and cdtB were detected by PCR. Molecular genotyping was performed by PCR ribotyping. Antimicrobial susceptibilities of the 120 C. difficile isolates were assessed by agar dilution methods. RESULTS Among 337 toxigenic isolates, 105 were toxin A-negative and toxin B-positive (A(-)B(+)) and 29 were binary toxin-producing strains. PCR ribotyping showed 50 different ribotype patterns. The 5 most frequently occurring ribotypes comprised 62.0% of all identified ribotypes. No isolate was susceptible to cefoxitin, and all except 1 were susceptible to piperacillin and piperacillin-tazobactam. The resistance rates of isolates to imipenem, cefotetan, moxifloxacin, ampicillin, and clindamycin were 25%, 34%, 42%, 51%, and 60%, respectively. The isolates showed no resistance to metronidazole or vancomycin. CONCLUSIONS This is the first nationwide study on the toxin status, including PCR ribotyping and antimicrobial resistance, of C. difficile isolates in Korea. The prevalence of A-B+ strains was 25.7%, much higher than that reported from other countries. Binary toxin-producing strains accounted for 7.1% of all strains, which was not rare in Korea. The most prevalent ribotype was ribotype 017, and all A-B+ strains showed this pattern. We did not isolate strains with decreased susceptibility to metronidazole or vancomycin.

Increasing prevalence and diversity of metallo-beta-lactamases in Pseudomonas spp., Acinetobacter spp., and Enterobacteriaceae from Korea.

ABSTRACT Among imipenem-nonsusceptible isolates, acquired metallo-β-lactamase genes were detected in 36 of 581 (6.2%) Pseudomonas aeruginosa isolates, 42 of 44 (95.4%) other Pseudomonas species, and 136 of 513 (26.5%) Acinetobacter species from 2003 to 2004 at a Korean hospital. Overall, blaVIM-2-like genes were the most prevalent and were also detected in Enterobacteriaceae, including Klebsiella pneumoniae.

Increase of Ceftazidime- and Fluoroquinolone-Resistant Klebsiella pneumoniae and Imipenem-Resistant Acinetobacter spp. in Korea: Analysis of KONSAR Study Data from 2005 and 2007

Purpose Antimicrobial resistance monitoring could be a useful source of information for treating and controlling nosocomial infections. We analyzed antimicrobial resistance data generated by Korean Hospitals and by a commercial laboratory in 2005 and 2007. Materials and Methods Susceptibility data for 2005 and 2007 were collected from 37 and 41 hospitals, respectively, and from one commercial laboratory. Intermediate susceptibility was not included in the calculation of resistance rates. Results Methicillin-resistant Staphylococcus aureus (MRSA) (64%), third-generation cephalosporin-resistant Klebsiella pneumoniae (29%), fluoroquinolone-resistant Escherichia coli (27%), Pseudomonas aeruginosa (33%), and Acinetobacter spp. (48%), and amikacin-resistant P. aeruginosa (19%) and Acinetobacter spp. (37%) were prevalent in hospitals in 2007. A gradual increase of vancomycin-resistant Enterococcus faecium and imipenem-resistant Acinetobacter spp. was observed. Higher incidences of third-generation cephalosporin-resistant E. coli and K. pneumoniae and imipenem-resistant P. aeruginosa were found in the commercial laboratory than in the hospitals. Conclusion Methicillin-resistant S. aureus, third-generation cephalosporin-resistant K. pneumoniae, and fluoroquinolone-resistant E. coli, P. aeruginosa and Acinetobacter spp. remain prevalent in Korea, while the incidence of vancomycin-resistant E. faecium and imipenem-resistant Acinetobacter spp. has increased gradually. The higher prevalences of third-generation cephalosporin-resistant E. coli and K. pneumoniae, and imipenem-resistant P. aeruginosa in the commercial laboratory are a new concern.

Necrotizing fasciitis involving the chest and abdominal wall caused by Raoultella planticola

BackgroundRaoultella planticola was originally considered to be a member of environmental Klebsiella. The clinical significance of R. planticola is still not well known.Case presentationWe describe the first case of necrotizing fasciitis involving the chest and abdominal wall caused by R. planticola. The identity of the organism was confirmed using 16S rRNA sequencing. The patient was successfully treated with the appropriate antibiotics combined with operative drainage and debridement.ConclusionsR. planticola had been described as environmental species, but should be suspected in extensive necrotizing fasciitis after minor trauma in mild to moderate immunocompromised patients.

New cfiA variant and novel insertion sequence elements in carbapenem-resistant Bacteroides fragilis isolates from Korea

Of 276 nonduplicate Bacteroides fragilis clinical isolates recovered from 1997 to 2004, 3 were resistant to carbapenem. cepA and cfiA alleles were detected by polymerase chain reaction in 240 (87.0%) and 11 (4.0%) of the isolates, respectively. Insertion sequence (IS) elements were found only in the 3 carbapenem-resistant B. fragilis isolates, which produced metallo-beta-lactamase at a level detectable by UV spectrophotometry. Sequence analysis showed 1 new cfiA variant, cfiA(11), and 2 novel IS elements. The cfiA(11) gene revealed 5 amino acid substitutions compared to cfiA, with 97.6% amino acid identity. The transposase, terminal inverted repeat sequence, and target site duplication sequence of the 2 novel IS elements were unique. This study reconfirmed the correlation between ISs and carbapenem resistance in B. fragilis.

First outbreak of multidrug-resistant Klebsiella pneumoniae producing both SHV-12-type extended-spectrum beta-lactamase and DHA-1-type AmpC beta-lactamase at a Korean hospital.

Purpose Coexistence of different classes of β-lactamases in a single bacterial isolate may pose diagnostic and therapeutic challenges. We investigated a spread of Klebsiella pneumoniae isolates co-producing an AmpC β-lactamase and an extended-spectrum β-lactamase (ESBL) in a university hospital. Materials and Methods Over a three-month period, a total of 11 K. pneumoniae isolates, which exhibited resistance to cefotaxime, aztreonam, and cefoxitin, were isolated. These isolates showed positive to ESBLs by double disk tests. Minimal inhibitory concentrations (MICs) were determined by broth microdilution testing. All isolates were examined by isoelectric focusing, PCR and sequence analysis to identify blaSHV and blaDHA, and molecular typing by pulsed-field gel electrophoresis (PFGE). Results All 11 isolates were highly resistant (MIC, ≥128 µg/ml) to ceftazidime, aztreonam, and cefoxitin, while they were susceptible (MIC, ≤2 µg/ml) to imipenem. The blaSHV-12 and blaDHA-1 genes were detected by PCR and sequence analysis. PFGE revealed a similar pattern in 10 of the 11 strains tested. Conclusion This is the first outbreak report of K. pneumoniae in Korea which co-produced SHV-12 and DHA-1 β-lactamase, and we suggest a clonal spread of multidrug-resistant K. pneumoniae at a hospital.

Evaluation of a rapid diagnostic test specific for Plasmodium vivax

Plasmodium vivax is the only human malaria indigenous to the Republic of Korea (ROK). A rapid and sensitive diagnostic test (RDT) that detects P. vivax is appropriate for evaluating suspected malaria patients with no travel history abroad. The RDTs, SD Malaria Antigen P.v (SD diagnostic, Kyonggi, ROK) specific for P. vivax and the well documented OptiMAL (DiaMed, Cressier, Switzerland) were compared among 282 volunteers for specificity and sensitivity of P. vivax and Plasmodium falciparum malaria infections against Giemsa‐stained blood smears read by an experienced microscopist. A total of 137 volunteers were diagnosed with P. vivax, 45 cases (returned travellers from overseas) were diagnosed with P. falciparum and 100 healthy volunteers were diagnosed as negative for malaria. Correspondingly, the SD Malaria Antigen P.v test identified P. vivax infections in 128/137 malaria patients (93.4%) and 0/100 (0%) healthy volunteers. Three patients identified with P. falciparum also were interpreted as P. vivax by the SD Malaria Antigen P.v test; however, these patients were later confirmed as mixed infections of P. vivax and P. falciparum by polymerase chain reaction. OptiMAL interpreted the three mixed infections only as P. falciparum and detected 130/137 (94.9%) patients with P. vivax. The sensitivity of the SD Malaria Antigen P.v test decreased from 100% (>5000 parasite/μl) to 81.3% (1–100 parasites/μl) as parasitaemia levels declined. For the regions where P. vivax is the primary malaria parasite, the SD P. vivax‐specific rapid diagnostic test may be useful for screening suspected malaria patients when sufficient material and human resources (e.g. trained microscopists) are unavailable for malaria diagnosis.

Prevalence of Inducible Clindamycin Resistance in Staphylococcal Isolates at a Korean Tertiary Care Hospital

Clindamycin resistance in Staphylococcus species can be either constitutive or inducible. Inducible resistance cannot be detected by the conventional antimicrobial susceptibility test. In this study, we determined the prevalence of inducible clindamycin resistance in staphylococcal isolates at a Korean tertiary care hospital. Between February and September 2004, 1,519 isolates of Staphylococcus aureus and 1,043 isolates of coagulase-negative staphylococci (CNS) were tested for inducible resistance by the D-zone test. Overall, 17% of MRSA, 84% of MSSA, 37% of MRCNS, and 70% of MSCNS were susceptible to clindamycin. Of the erythromycin non-susceptible, clindamycin-susceptible isolates, 32% of MRSA, 35% of MSSA, 90% of MRCNS, and 94% of MSCNS had inducible clindamycin resistance. Inducible clindamycin resistance in staphylococci was highly prevalent in Korea. This study indicates importance of the D-zone test in detecting inducible clindamycin resistance in staphylococci to aid in the optimal treatment of patients.

Isolation of a Klebsiella pneumoniae Isolate of Sequence Type 258 Producing KPC-2 Carbapenemase in Korea

Carbapenem-resistant Klebsiella pneumoniae isolates producing K. pneumoniae carbapenemases (KPC) were first reported in the USA in 2001, and since then, this infection has been reported in Europe, Israel, South America, and China. In Korea, the first KPC-2-producing K. pneumoniae sequence type (ST) 11 strain was detected in 2010. We report the case of a patient with a urinary tract infection caused by KPC-2-producing K. pneumoniae. This is the second report of a KPC-2-producing K. pneumoniae infection in Korea, but the multilocus sequence type was ST258. The KPC-2-producing isolate was resistant to all tested β-lactams (including imipenem and meropenem), amikacin, tobramycin, ciprofloxacin, levofloxacin, and trimethoprim-sulfamethoxazole, but was susceptible to gentamicin, colistin, polymyxin B, and tigecycline. The KPC-2-producing isolate was negative to phenotypic extended-spectrum β-lactamase (ESBL) and AmpC detection tests and positive to modified Hodge test and carbapenemase inhibition test with aminophenylboronic acid.

Evaluation of BACTEC Plus aerobic and anaerobic blood culture bottles and BacT/Alert FAN aerobic and anaerobic blood culture bottles for the detection of bacteremia in ICU patients

Blood culture is the most valuable laboratory test for the diagnosis of bacteremia and sepsis. The BACTEC FX and BacT/Alert 3D automated blood culture systems are commonly used in Korean health care facilities. A controlled clinical evaluation of the resin-containing BACTEC Plus aerobic (BA) and anaerobic (BN), and the charcoal-containing FAN aerobic (FA) and anaerobic (FN) bottles using blood from intensive care unit (ICU) patients was designed. The performances of these 2 systems with media containing particle absorbing antimicrobial agents were evaluated using the culture positivity rate and time to detection (TTD). TTD was collected using data management systems, either the Epicenter (BD Diagnostic Systems) or the hospital laboratory information system. A total of 1539 four-bottle sets were collected from 270 patients in medical and surgical ICUs. Blood culture samples included 1539 bottles each of BA, BN, FA, and FN, and yielded 113 (7.3%), 90 (5.8%), 104 (6.8%), and 80 (5.2%) positive bacterial or fungal isolates, respectively. There were significant differences between the resin-containing BA and BN samples in culture positivity and also between the charcoal-containing FA and FN samples, especially for Escherichia coli (25/27 versus 17/27, P < 0.05) and Acinetobacter baumannii (14/15 versus 7/15, P < 0.05). Significantly shorter recovery time was observed in BACTEC Plus aerobic bottles than in FAN aerobic bottles (17.2 and 24.7 h, respectively) (P < 0.001).