An Effective Method of RNA Extraction from Mycobacterium tuberculosis / 대한임상미생물학회지

In the RNA-based study, it is important to extract high-quality RNA. However, RNA extraction from Mycobacterium tuberculosis is problematic due to its thick, waxy cell wall rich in mycolic acid, which renders the cells resistant to lysis. Using TRIzol reagent and several powerful bead-beating steps, a high quantity of RNA was obtained.

Trends in Bloodstream Infections at a Korean University Hospital between 2008 and 2013 / 대한임상미생물학회지

BACKGROUND: Blood culture remains the definitive method for diagnosing bacteremia and fungemia. In this study, we investigated the incidence of bacterial and fungal infections along with the trends in antimicrobial susceptibility in blood cultures collected from 2008 to 2013. METHODS: We performed a retrospective analysis of blood cultures performed at Kyung Hee University Hospital, Seoul, South Korea, between 2008 and 2013 to determine the bacterial and fungal species isolated, and their antimicrobial susceptibilities. Additional analyses were performed comparing these results to that of a prior study examining blood cultures collected from 2003-2007. RESULTS: Of the 102,257 specimens collected, 8,452 (8.3%) were culture positive, with Staphylococcus epidermidis being the most common species isolated (17.3%), followed by Escherichia coli (16.9%), Staphylococcus aureus (8.1%), and Klebsiella pneumoniae (6.5%). Fungal species accounted for 3.7% of all isolates. Methicillin resistance was seen in 54.3% of S. aureus isolates. The frequencies of extended-spectrum beta-lactamase (ESBL)-producing E. coli and K. pneumoniae were 13.1% and 10.3%; imipenem resistance was seen in 19.5% of Pseudomonas aeruginosa isolates. CONCLUSION: Although the number of blood specimens analyzed increased steadily over the course of this study, the rate of positive blood cultures declined. The most common microorganisms isolated were coagulase-negative staphylococci, E. coli, S. aureus, and K. pneumoniae, consistent with our prior analysis. This analysis of blood culture isolate frequencies and antibiotic susceptibilities can be used to inform antibiotic therapy decisions.

A Study of Efflux Pump Genes in Mycobacterium tuberculosis Clinical Isolates / 대한임상미생물학회지

The efflux pump system has been suggested as an important mechanism in the drug resistance of Mycobacterium tuberculosis (MTB). In this study, molecular analysis of five genes in the efflux pump system of MTB isolates from Korean patients was performed in order to identify appropriate molecular targets. In this study, 35 culture-positive specimens were included. PCR was performed for five efflux genes, mmpL7, efpA, mmr, p55 and tap-like gene. In the 35 clinical isolates, molecular analysis of five kinds of efflux pump genes was performed. Only one clinical isolate showed negative PCR results for all five efflux pump genes. All the rest 34 isolates presented concurrent positive results for the five efflux pump genes. In the near future, gene expression study with quantitative PCR should be performed using these genes.

Evaluation of a Chromogenic Culture Medium for the Detection of Clostridium difficile

PURPOSE: Clostridium difficile (C. difficile) is an important cause of nosocomial diarrhea. Diagnostic methods for detection of C. difficile infection (CDI) are shifting to molecular techniques, which are faster and more sensitive than conventional methods. Although recent advances in these methods have been made in terms of their cost-benefit, ease of use, and turnaround time, anaerobic culture remains an important method for detection of CDI. MATERIALS AND METHODS: In efforts to evaluate a novel chromogenic medium for the detection of C. difficile (chromID CD agar), 289 fecal specimens were analyzed using two other culture media of blood agar and cycloserine-cefoxitin-fructose-egg yolk agar while enzyme immunosorbent assay and polymerase chain reaction-based assay were used for toxin detection. RESULTS: ChromID showed the highest detection rate among the three culture media. Both positive rate and sensitivity were higher from chromID than other culture media. ChromID was better at detecting toxin producing C. difficile at 24 h and showed the highest detection rate at both 24 h and 48 h. CONCLUSION: Simultaneous use of toxin assay and anaerobic culture has been considered as the most accurate and sensitive diagnostic approach of CDI. Utilization of a more rapid and sensitive chromogenic medium will aid in the dianogsis of CDI.

16S Ribosomal RNA Identification of Prevotella nigrescens from a Case of Cellulitis

No abstract available.

Quinolone-resistant Shigella flexneri Isolated in a Patient Who Travelled to India

We report a recent case in which ciprofloxacin-resistant Shigella flexneri was isolated from a 23-yr-old female patient with a history of travel to India. Prior to her admission to our internal medicine department, she experienced symptoms of high fever and generalized weakness from continuous watery diarrhea that developed midway during the trip. S. flexneri was isolated from the stool culture. Despite initial treatment with ciprofloxacin, the stool cultures continued to show S. flexneri growth. In the susceptibility test for antibiotics of the quinolone family, the isolate showed resistance to ciprofloxacin (minimum inhibitory concentration [MIC], 8 microg/mL), norfloxacin (MIC, 32 microg/mL), ofloxacin (MIC, 8 microg/mL), nalidixic acid (MIC, 256 microg/mL), and intermediate resistance to levofloxacin (MIC, 4 microg/mL). In molecular studies for quinolone resistance related genes, plasmid borne-quinolone resistance genes such as qnrA, qnrB, qnrS, aac(6')-Ib-cr, qepA, and oqxAB were not detected. Two mutations were observed in gyrA (248C-->T, 259G-->A) and 1 mutation in parC (239G-->T). The molecular characteristics of the isolated S. flexneri showed that the isolate was more similar to the strains isolated from the dysentery outbreak in India than those isolated from Korea.

Characteristics of Outpatients with Pandemic H1N1/09 Influenza in a Tertiary Care University Hospital in Korea

The pandemic H1N1/09 emerged rapidly in Korea. Here, we describe the clinical characteristics of outpatients in Seoul, Korea who were infected in the 2009 H1N1 pandemic. We reviewed the cases of outpatients with pandemic H1N1/09 who visited a tertiary care teaching hospital between September 1 and December 31, 2009. Infection with pandemic H1N1/09 was confirmed by molecular tests. Of a total of 7,182 tests, 3,020 (42.0%) were positive. Compared with 473 cases of influenza-like illness (ILI), the 586 confirmed cases of pandemic H1N1/09 differed in age [odds ratio (OR) 0.975] and fulfilling at least one of the following factors: age or =65 years, history of contact with other pandemic H1N1/09-infected individuals (OR 0.611), fever > or =37.8degrees C (OR 3.567), cough (OR 2.290), and myalgia (OR 1.559). The sensitivity of the best criteria, "fever (> or =37.8degrees C) plus cough" (41.03%) in this study was lower than that of the Korea Centers for Disease Control and Prevention (KCDC) criteria (47.95%), whereas the positive likelihood ratio (3.55) and positive predictive value (81.6) of this criteria was higher than those of the KCDC criteria (2.98 and 78.7, respectively). The clinical characteristics of pandemic H1N1/09 are, in many regards, indistinguishable from those of ILI. Moreover, the accuracy and predictability of criteria which include only symptoms or signs were not sufficient to diagnose pandemic H1N1/09 infection. Therefore, use of a combination of symptoms with confirmatory laboratory testing is necessary for accurate diagnosis of pandemic H1N1/09.