Kyu Man Lee

Microbiologic aspects of predominant bacteria isolated from the burn patients in Korea

The risk of infection in burns is well-known. In recent decades, the antimicrobial resistance of bacteria isolated from burn patients has increased. For this reason, we have carried out a study of the predominant bacterial profiles and antimicrobial resistance patterns of isolates from a burn center in Korea. A retrospective study was undertaken at Hallym University, Hangang Sacred Heart Hospital to examine the bacterial isolates from the burn patients and to compare the antibiograms of the predominant bacteria isolated from these patients with those of the other wards over a period of 3 years. Pseudomonas aeruginosa was the most common (n=2997, 45.7%) isolate from the burn patients followed by Staphylococcus aureus (n=21261, 19.2%) and Acinetobacter baumannii (n=878, 13.4%). These bacteria, isolated from the burn patients, were almost all higher in antimicrobial resistance rate than those in the non-burn patients (P<0.05). Because these bacteria showed very high resistant rates, they must be avoided in order to control a hospital-acquired infection. Our results seem helpful in providing useful guidelines for choosing effective empiric antimicrobial therapy against bacteria isolated from the burn patients in Korea.

Boronic acid disk tests for identification of extended-spectrum β-lactamase production in clinical isolates of Enterobacteriaceae producing chromosomal AmpC β-lactamases

A study using boronic acid (BA) was designed to detect the extended-spectrum beta-lactamases (ESBLs) in Enterobacteriaceae producing chromosomal AmpC beta-lactamases. A total of 197 clinical isolates of Enterobacter spp. (n=100), Serratia marcescens (n=62) and Citrobacter freundii (n=35) were analysed. Genes encoding ESBLs were detected by polymerase chain reaction (PCR) amplification followed by direct sequencing of PCR products. The Clinical and Laboratory Standards Institute confirmatory test detected only 72.1% of the ESBL-producing isolates. When a > or =5mm increase in the zone diameter of either the cefotaxime/clavulanic acid and/or the ceftazidime/clavulanic acid disks tested in combination with BA versus cefotaxime and/or ceftazidime containing BA was considered to be a positive for ESBL, the method detected 60 (98.4%) of the 61 isolates that harboured ESBLs and showed no false-positive results for ESBL-non-producing isolates. In conclusion, the BA disk test is a highly sensitive and specific method for the detection of ESBLs in Enterobacteriaceae producing chromosomal AmpC beta-lactamases.

Comparison of Diagnostic Utility between Procalcitonin and C-Reactive Protein for the Patients with Blood Culture-Positive Sepsis

BACKGROUND Procalcitonin (PCT) is a relatively new marker for bacterial infections, and its diagnostic utility has been variable across the studies. We investigated the diagnostic utility of PCT for the patients with blood culture-positive sepsis, and compared it with that of C-reactive protein (CRP). METHODS In 1,270 consecutive blood samples, PCT and CRP were simultaneously measured and results were compared according to the five categories of PCT concentrations (<0.05 ng/mL; 0.05-0.49 ng/mL; 0.5-1.99 ng/mL; 2-9.99 ng/mL; > or = 10 ng/mL). In 506 samples, they were further analyzed according to the result of blood culture. PCT and CRP were measured using enzyme-linked fluorescent assay (bioMerieux Co., France) and rate nephelometry (Beckman Coulter Co., USA), respectively. Their diagnostic utilities were compared using ROC curves. RESULTS The mean concentrations of CRP in five categories of PCT were 15.4 mg/L, 42.1 mg/L, 101.2 mg/L, 125.0 mg/L, 167.1 mg/L, respectively (P<0.0001). Both PCT and CRP showed significant differences between the two positive and negative groups of blood culture (PCT, 8.47 vs 2.44 ng/mL, P=0.0133; CRP, 110.48 vs 59.78 mg/L, P<0.0001). The areas under the ROC curves (95% confidence interval) for PCT and CRP were 0.720 (0.644-0.788) and 0.558 (0.478-0.636), respectively, and showed a significant difference (P=0.005). CONCLUSIONS The diagnostic utility of PCT is superior to that of CRP for the patients with blood culture-positive sepsis. PCT seems to be reliable for sepsis diagnosis, and may provide useful information for the critically ill patients.

Clonal and horizontal spread of the blaOXA-232 gene among Enterobacteriaceae in a Korean hospital

All 16 Klebsiella pneumoniae isolates and both Escherichia coli isolates harbored the bla(OXA-232) and bla(CTX-M-15) genes. Furthermore, all 16 K. pneumoniae isolates belonged to a unique pulsed-field gel electrophoresis clone and were assigned to an identical sequence type (ST14). The 2 E. coli isolates were identified as ST131 and ST457. The bla(OXA-232) gene underwent horizontal transfer to E. coli isolates via a conjugative ColE-type plasmid. The introduction of this K. pneumoniae ST14 strain to the Korean hospital was attributed to an index patient who was likely colonized during a prior hospitalization in India.

Contamination of X-ray cassettes with methicillin resistant Staphylococcus aureus and methicillin-resistant Staphylococcus haemolyticus in a radiology department.

Background We performed surveillance cultures of the surfaces of X-ray cassettes to assess contamination with methicillin-resistant Staphylococcus aureus (MRSA). Methods The surfaces of 37 X-ray cassettes stored in a radiology department were cultured using mannitol salt agar containing 6 µg/mL oxacillin. Suspected methicillin-resistant staphylococcal colonies were isolated and identified by biochemical testing. Pulsed-field gel electrophoresis (PFGE) analysis was performed to determine the clonal relationships of the contaminants. Results Six X-ray cassettes (16.2%) were contaminated with MRSA. During the isolation procedure, we also detected 19 X-ray cassettes (51.4%) contaminated with methicillin-resistant Staphylococcus haemolyticus (MRSH), identified as yellow colonies resembling MRSA on mannitol salt agar. PFGE analysis of the MRSA and MRSH isolates revealed that most isolates of each organism were identical or closely related to each other, suggesting a common source of contamination. Conclusions X-ray cassettes, which are commonly in direct contact with patients, were contaminated with MRSA and MRSH. In hospital environments, contaminated X-ray cassettes may serve as fomites for methicillin-resistant staphylococci.

Evaluation of the SD Bioline Norovirus rapid immunochromatography test using fecal specimens from Korean gastroenteritis patients

Abstract The analytical and clinical performance of a new rapid immunochromatography test, the SD Bioline Norovirus test, was evaluated for the detection of human norovirus in fecal specimens. The analytical performance studies were performed for detection limit, reproducibility, cross-reactivity, and interference. For comparison, 92 norovirus-positive stool samples and 126 norovirus-negative samples for which the results were confirmed by 2 different real-time PCR kits were used. The rapid immunochromatography test detected the equivalent of 4.48×106 copies/mL of the norovirus genome in stool samples. On performing the repeatability/reproducibility test, samples above this concentration all provided positive results (100%) and 97.8% of the samples slightly below this concentration (2.45×106 copies/mL) provided negative results. No cross-reactivity or interference was detected. Positive percent agreement (sensitivity), negative percent agreement (specificity), and overall percent agreement of the rapid immunochromatography test compared with testing by real-time PCR were 90.2%, 100%, and 95.9%, respectively. In addition, the rapid immunochromatography test was completed within 20min. The SD Bioline Norovirus test was, therefore, easier and more rapid to perform and showed excellent reproducibility, no cross-reactivity, no interference, and high agreement compared with real-time PCR. Thus, this test is useful for rapid screening to identity norovirus infection.

Epidemiological Analysis of Norovirus Infection between March 2007 and February 2010

BACKGROUND Norovirus is a common cause of non-bacterial acute gastroenteritis worldwide, and norovirus infection shows symptoms such as vomiting and diarrhea in patients of all age groups. Mass outbreaks of norovirus infection have been recently reported in Korea. Herein, we investigated the epidemiological characteristics of acute norovirus gastroenteritis. METHODS We analyzed 11,219 fecal specimens of patients with acute gastroenteritis symptoms from the 5 participating hospitals for 3 yr (March 2007-February 2010) to determine positive rates of detection using RIDASCREEN Norovirus ELISA (R-Biopharm AG, Germany) kit by year, prevalence season, sex, age, and region. RESULTS Norovirus infection was prevalent during autumn and winter, and 13.0% specimens were positive for the infection. The positive rates of norovirus detection were 16.2%, 13.8%, and 9.9% in 2007, 2008, and 2009, respectively, and they tended to decrease every year. In 2007 and 2008, the epidemicity of norovirus started from October, reached its peak in November, and lasted until January. However, in 2009, it started from December, reached its peak in January, and lasted until February. Most patients were 0-3 yr old and this patient group had the highest positive rate. There was no significant inter-regional difference among the subjects. CONCLUSIONS We performed epidemiological analysis of norovirus infection using ELISA assay. Reverse transcription-PCR indicated higher prevalence of norovirus infection as compared with that reported before 2007. Further studies are warranted to examine the changes observed in the epidemic period of 2009.

Evaluation of an immunochromatographic assay for the rapid and simultaneous detection of rotavirus and adenovirus in stool samples.

Background We evaluated the analytical and clinical performances of the SD BIOLINE Rota/Adeno Rapid kit (SD Rota/Adeno Rapid; Standard Diagnostics, Inc., Korea), an immunochromatographic assay (ICA), for the simultaneous detection of rotaviruses and adenoviruses in human stool samples. Methods We tested 400 clinical stool samples from patients with acute gastroenteritis and compared the ICA results with the results obtained by using ELISA, enzyme-linked fluorescent assays (ELFA), PCR, and multiplex reverse transcription-PCR (mRT-PCR). To assess the analytical performance of the SD BIOLINE Rota/Adeno Rapid kit, we determined its detection limit, reproducibility, cross-reactivity, and analytical reactivity for adenovirus subtypes, and performed interference studies. Results The overall agreement rates among the tested methods were 91.5% for rotavirus and 85.5% for adenovirus. On the basis of mRT-PCR, the overall agreement, positive agreement, and negative agreement rates of the ICA were 95.6%, 100%, and 94.9% for rotavirus, and 94.0%, 71.4%, and 94.8% for adenovirus, respectively. Using the ICA, we detected all the subtypes of adenovirus tested, but the analytical reactivities for adenovirus subtypes were different between the 4 adenovirus detection methods. The high reproducibility was confirmed, and no cross-reactivity or interference was detected. Conclusions The SD BIOLINE Rota/Adeno Rapid kit showed acceptable analytical and clinical performances. However, interpretation of adenovirus positive/negative result should be cautious because of different detectability for adenovirus subtypes among adenovirus detection methods.

Broth Microdilution Method To Detect Extended-Spectrum β-Lactamases and AmpC β-Lactamases in Enterobacteriaceae Isolates by Use of Clavulanic Acid and Boronic Acid as Inhibitors

ABSTRACT This study was designed to evaluate the performance of the broth microdilution (BMD) method to detect production of extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases in Enterobacteriaceae by using clavulanic acid (CA) and boronic acid (BA) as ESBL and AmpC β-lactamase inhibitors, respectively. A total of 100 clinical isolates of Enterobacteriaceae were analyzed. Mueller-Hinton broth containing serial twofold dilutions of cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), or cefepime (FEP) with or without either or both CA and BA was prepared. An eightfold or greater decrease in the MIC of CTX, CAZ, ATM, or FEP in the presence of CA and BA was considered a positive result for ESBL and plasmid-mediated AmpC β-lactamase (PABL), respectively. In tests with CA, expanded-spectrum β-lactams containing BA (CTX-BA, CAZ-BA, ATM-BA, and FEP-BA) showed higher positive rates in detecting ESBL producers than those without BA. The combination of CTX- and CAZ-based BMD tests with CA and BA showed sensitivity and specificity of 100% for the detection of ESBLs and PABLs. The BMD testing could be applicable for routine use in commercially available semiautomated systems for the detection of ESBLs and PABLs in Enterobacteriaceae.

Evaluation of a new real-time reverse transcription polymerase chain reaction assay for detection of norovirus in fecal specimens

A new real-time reverse transcription polymerase chain reaction (RT-PCR) assay, the AccuPower Norovirus Real-time RT-PCR Kit, was evaluated in detection of human norovirus in stool specimens. Studies for detection limit, dynamic range, reproducibility, and cross-reactivity were performed. A total of 281 fecal specimens were tested using the AccuPower Norovirus Real-time RT-PCR Kit, and the results were compared with those obtained using another real-time RT-PCR system. Norovirus positivity and genotype were confirmed by direct sequencing. The lowest mean numbers of genome copies of GI and GII that could be detected by the assay were 12.3 and 5.6 RNA copies/reaction, respectively. The positive, negative, and overall percent agreements between the 2 real-time PCR assays were 99.0% (96/97), 95.1% (175/184), and 96.4% (271/281), respectively. The AccuPower Norovirus Real-time RT-PCR system showed good analytical and clinical performance and may be a useful diagnostic tool for norovirus infection.