Myungsook Kim

Increasing Prevalence of Toxin A-Negative, Toxin B-Positive Isolates of Clostridium difficile in Korea: Impact on Laboratory Diagnosis

ABSTRACT Of 462 Korean Clostridium difficile isolates, 77.5% were toxin B positive but 21.4% were toxin A negative (A− B+). The binary toxin gene was detected in nine isolates. A higher fluoroquinolone resistance of A− B+ strains may contribute to the increase of these strains. Toxin A detection alone may underdiagnose C. difficile-associated disease.

Evaluation of the Xpert Clostridium difficile Assay for the Diagnosis of Clostridium difficile Infection

Infection with Clostridium difficile is a growing concern because of the increasing prevalence and spread of nosocomial infections. Emergence of the hypervirulent 027/NAP1/BI strain is also notable. Existing diagnostic methods have low sensitivity or are time-consuming. Therefore, establishing a rapid and accurate microbiological diagnostic assay is needed. We evaluated the Xpert C. difficile assay (Xpert CD assay; Cepheid, USA) to detect toxigenic C. difficile. This assay is a real-time multiplex PCR assay that can be used to detect toxigenic C. difficile strains and differentiate the C. difficile presumptive 027/NAP1/BI strain. A total of 253 loose stool specimens were collected and toxigenic cultures, VIDAS C. difficile A & B assays (VIDAS CDAB assay; bioMérieux, France), and the Xpert CD assay were performed. In comparison to toxigenic cultures, the sensitivity, specificity, and positive and negative predictive values were 100%, 94.6%, 83.1%, and 100%, respectively, for the Xpert CD assay and 40.8%, 98.0%, 100%, and 88.9%, respectively, for VIDAS CDAB assay. Because of the low prevalence of the PCR ribotype 027 in Korea, the evaluation of the usefulness of the Xpert CD assay for screening for the 027 strain was limited. The Xpert CD assay provides great sensitivity in diagnosing toxigenic C. difficile infection. In addition, this method has excellent usability because it is simple and fast.

Relative prevalence and antimicrobial susceptibility of clinical isolates of Elizabethkingia species based on 16S rRNA gene sequencing.

ABSTRACT Some of the previously reported clinical isolates of Elizabethkingia meningoseptica may be later named species of Elizabethkingia. We determined the accuracy of species identification (with two matrix-assisted laser desorption ionization–time of flight mass spectrometry [MALDI-TOF MS] systems and the Vitek 2 GN card), relative prevalence of three Elizabethkingia spp. in clinical specimens, and antimicrobial susceptibility of the species identified by 16S rRNA gene sequencing. Specimens for culture were collected from patients in a university hospital in Seoul, South Korea, between 2009 and 2015. All 3 Elizabethkingia spp. were detected in patients; among the 86 isolates identified by 16S rRNA gene sequencing, 17 (19.8%) were E. meningoseptica, 18 (20.9%) were Elizabethkingiamiricola, and 51 (59.3%) were Elizabethkingiaanophelis. Only the MALDI-TOF Vitek MS system with an amended database correctly identified all of the isolates. The majority (76.7%) of the isolates were from the lower respiratory tract, and 8 (9.3%) were from blood. Over 90% of E. meningoseptica and E. anophelis isolates were susceptible to piperacillin-tazobactam and rifampin. In contrast, all E. miricola isolates were susceptible to fluoroquinolones except ciprofloxacin. Further studies are urgently needed to determine the optimal antimicrobial agents for the treatment of infections due to each individual Elizabethkingia species.

Evaluation of VITEK mass spectrometry (MS), a matrix-assisted laser desorption ionization time-of-flight MS system for identification of anaerobic bacteria.

Background By conventional methods, the identification of anaerobic bacteria is more time consuming and requires more expertise than the identification of aerobic bacteria. Although the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems are relatively less studied, they have been reported to be a promising method for the identification of anaerobes. We evaluated the performance of the VITEK MS in vitro diagnostic (IVD; 1.1 database; bioMérieux, France) in the identification of anaerobes. Methods We used 274 anaerobic bacteria isolated from various clinical specimens. The results for the identification of the bacteria by VITEK MS were compared to those obtained by phenotypic methods and 16S rRNA gene sequencing. Results Among the 249 isolates included in the IVD database, the VITEK MS correctly identified 209 (83.9%) isolates to the species level and an additional 18 (7.2%) at the genus level. In particular, the VITEK MS correctly identified clinically relevant and frequently isolated anaerobic bacteria to the species level. The remaining 22 isolates (8.8%) were either not identified or misidentified. The VITEK MS could not identify the 25 isolates absent from the IVD database to the species level. Conclusions The VITEK MS showed reliable identifications for clinically relevant anaerobic bacteria.

First Report of Brain Abscess Associated with Pseudozyma species in a Patient with Astrocytoma

A yeast-like strain was isolated from the brain abscess of a patient diagnosed with astrocytoma. Morphological and molecular analysis on D1/D2 domain in the 26S rRNA gene and internal transcript spacer region of the strain revealed that the strain belonged to the genus Pseudozyma. To the best of our knowledge, this is the first report on the isolation of a Pseudozyma strain from brain abscess.

Antimicrobial susceptibility of Stenotrophomonas maltophilia isolates from a Korean tertiary care hospital

We determined the antimicrobial susceptibility of 90 clinical isolates of Stenotrophomonas maltophilia collected in 2009 at a tertiary care hospital in Korea. Trimethoprim-sulfamethoxazole, minocycline, and levofloxacin were active against most of the isolates tested. Moxifloxacin and tigecycline were also active and hold promise as therapeutic options for S. maltophilia infections.

Insufficient Discriminatory Power of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Dendrograms to Determine the Clonality of Multi-Drug-Resistant Acinetobacter baumannii Isolates from an Intensive Care Unit

While pulsed-field gel electrophoresis (PFGE) is recognized as the gold standard method for clonality analysis, MALDI-TOF MS has recently been spotlighted as an alternative tool for species identification. Herein, we compared the dendrograms of multi-drug-resistant (MDR) Acinetobacter baumannii isolates by using MALDI-TOF MS with those by using PFGE. We used direct colony and protein extraction methods for MALDI-TOF MS dendrograms. The isolates with identical PFGE patterns were grouped into different branches in MALDI-TOF MS dendrograms. Among the isolates that were classified as very close isolates in MALDI-TOF MS dendrogram, PFGE band patterns visually showed complete differences. We numeralized similarity among isolates by measuring distance levels. The Spearman rank correlation coefficient values were 0.449 and 0.297 between MALDI-TOF MS dendrogram using direct colony and protein extraction method versus PFGE, respectively. This study is the first paper focusing solely on the dendrogram function of MALDI-TOF MS compared with PFGE. Although MALDI-TOF MS is a promising tool to identify species in a rapid manner, our results showed that MALDI-TOF MS dendrograms could not substitute PFGE for MDR Acinetobacter baumannii clonality analysis.

Mixed finite element domain decomposition for nonlinear parabolic problems

Abstract Fully discrete mixed finite element method is considered to approximate the solution of a nonlinear second-order parabolic problem. A massively parallel iterative procedure based on domain decomposition technique is presented to solve resulting nonlinear algebraic equations. Robin type boundary conditions are used to transmit information between subdomains. The convergence of the iteration for each time step is demonstrated. Optimal-order error estimates are also derived. Numerical examples are given.

Anaerobic Bacteremia: Impact of Inappropriate Therapy on Mortality

Background Investigation on incidence and mortality of anaerobic bacteremia (AB) is clinically relevant in spite of its infrequent occurrence and not often explored, which report varies according to period and institutions. Therefore, it is necessary to analyze the incidence and risk factors related to mortality and assess clinical outcomes of AB in current aspect. Materials and Methods Characteristics of AB patients and anaerobic bacteria from blood culture at a university hospital in 2012 were reviewed retrospectively. The correlation between risk factors and 28-day patient mortality was analyzed. Results A total of 70 non-duplicated anaerobic bacteria were isolated from blood of 70 bacteremia patients in 2012. The history of cardiovascular disease as hosts risk factor was statistically significant (P = 0.0344) in univariate and multivariate analysis. Although the inappropriate therapy was not statistically significant in univariate and multivariate analysis, the survival rate of bacteremia was significantly worse in patients who had inappropriate therapy compared with those underwent appropriate therapy (hazard ratio, 5.4; 95% confidence interval, 1.7–6.9; P = 0.004). The most frequently isolated organism was Bacteroides fragilis (32 isolates, 46%), followed by Bacteroides thetaiotaomicron (10, 14%), and non-perfringens Clostridium (7, 10%). Conclusion The incidence of AB in 2012 was 2.3% (number of AB patients per 100 positive blood culture patients) and the mortality rate in patients with clinically significant AB was 21.4%. In addition, AB was frequently noted in patients having malignancy and the survival rate of AB was significantly worse in patients who received inappropriate therapy compared with those underwent appropriate therapy.

Characterization of the Multidrug-Resistant Acinetobacter species Causing a Nosocomial Outbreak at Intensive Care Units in a Korean Teaching Hospital: Suggesting the Correlations with the Clinical and Environmental Samples, Including Respiratory Tract-related Instruments

Background: Acinetobacter spp. is an important nosocomial pathogen for which increasing resistance to multiple antimicrobial agents has been observed. Prevalence of multidrug-resistant (MDR) Acinetobacter spp. in the intensive care unit (ICU) at a teaching hospital in Korea started to increase in 2008. The aim of this study was to determine the source of pathogen spread and to characterize the emerging strains at an early stage of outbreak. Methods: Samples from respiratory instruments and fomites in the ICUs, as well as from the healthcare workers, were cultured to identify the sources of MDR Acinetobacter spp. Antimicrobial susceptibility was determined by the CLSI disk diffusion method. Pulsed field gel electrophoresis (PFGE) was performed for clinical and environmental isolates in order to determine clonality. Carbapenemase genes were detected by multiplex PCR. Infection control measures including peer-monitoring of hand washing, environmental cleaning and standard precautions were enforced. Results: Among the samples from the ICU tools (105) and healthcare worker’s hands (44), 31 (30%) and 2 (5%) respective samples yielded MDR Acinetobacter spp. Among the environmental samples, 90% were from respiratory-related equipment. The majority of clinical and environmental MDR Acinetobacter spp. (44/55) belonged to the pulsotype A. baumannii and carried both blaOXA-51-like and blaOXA-23-like genes. Even though infection-control measures were enforced, prevalence of MDR Acinetobacter spp. continues to increase. Conclusion: An outbreak of MDR Acinetobacter spp. in a Korean hospital was caused by A. baumannii carrying the blaOXA-23-gene and was correlated with contaminated respiratory-related instruments in the ICUs. More intensive measures for nosocomial infection control are needed for successful prevention of Acinetobacter spread in hospitals. (Ann Clin Microbiol 2014;17:29-34)