BACKGROUND: The aims of this study were to compare several DNA extraction methods and 16S rDNA primers and to evaluate the clinical utility of broad-range PCR in continuous ambulatory peritoneal dialysis (CAPD) culture fluids. METHODS: Six type strains were used as model organisms in dilutions from 10(8) to 100 colony-forming units (CFU)/mL for the evaluation of 5 DNA extraction methods and 5 PCR primer pairs. Broad-range PCR was applied to 100 CAPD culture fluids, and the results were compared with conventional culture results. RESULTS: There were some differences between the various DNA extraction methods and primer sets with regard to the detection limits. The InstaGene Matrix (Bio-Rad Laboratories, USA) and Exgene Clinic SV kits (GeneAll Biotechnology Co. Ltd, Korea) seem to have higher sensitivities than the others. The results of broad-range PCR were concordant with the results from culture in 97% of all cases (97/100). Two culture-positive cases that were broad-range PCR-negative were identified as Candida albicans, and 1 PCR-positive but culture-negative sample was identified as Bacillus circulans by sequencing. Two samples among 54 broad-range PCR-positive products could not be sequenced. CONCLUSIONS: There were differences in the analytical sensitivity of various DNA extraction methods and primers for broad-range PCR. The broad-range PCR assay can be used to detect bacterial pathogens in CAPD culture fluid as a supplement to culture methods.
Bacillus/genetics/isolation & purification ; Bacteria/genetics/*isolation & purification ; Candida albicans/genetics/isolation & purification ; DNA Primers/genetics ; DNA, Bacterial/*analysis/isolation & purification ; *Genetic Techniques/standards ; Humans ; Peritoneal Dialysis, Continuous Ambulatory ; Peritonitis/*microbiology ; Polymerase Chain Reaction ; Reagent Kits, Diagnostic ; Sequence Analysis, DNA

BACKGROUND: We investigated the prevalence of plasmid-mediated quinolone resistance and its association with extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase in Enterobacteriaceae. METHODS: A total of 347 non-duplicated isolates of Enterobacteriaceae were collected between August and October 2006 from 2 hospitals. Qnr determinant screening was conducted using PCR amplification, and all positive results were confirmed by direct sequencing. Qnr-positive strains were determined on the basis of the presence of ESBL and AmpC beta-lactamase genes. RESULTS: The qnr gene was detected in 47 of 347 clinical Enterobacteriaceae isolates. Among the 47 qnr-positive strains, Klebsiella pneumoniae (N=29) was the most common, followed by Escherichia coli (N=6), Enterobacter cloacae (N=6), Citrobacter freundii (N=5), and Enterobacter aerogenes (N=1). These isolates were identified as qnrA1 (N=6), 8 qnrB subtypes (N=40), and qnrS1 (N=1). At least 1 ESBL was detected in 38 of the 47 qnr-positive strains. Qnr-positive strains also showed high positive rates of ESBL or AmpC beta-lactamase, such as TEM, SHV, CTX-M, and DHA. DHA-1 was detected in 23 of 47 qnr-positive strains, and this was co-produced with 1 qnrA1 and 22 qnrB4. Strains harboring MIR-1T and CMY were also detected among the qnr-positive strains. Antimicrobial-resistance rates of qnr-positive strains to ciprofloxacin, levofloxacin, norfloxacin, nalidixic acid, and moxifloxacin were 51.1%, 46.8%, 46.8%, 74.5%, and 53.2%, respectively. CONCLUSIONS: The qnr genes were highly prevalent in Enterobacteriaceae, primarily the qnrB subtypes. They were closely associated with EBSL and AmpC beta-lactamase.
Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/biosynthesis/*genetics ; DNA, Bacterial/chemistry/genetics ; Drug Resistance, Bacterial/*genetics ; Enterobacteriaceae/enzymology/*genetics/isolation & purification ; Enterobacteriaceae Infections/microbiology ; *Genetic Variation ; Hospitals, University ; Humans ; Microbial Sensitivity Tests ; Plasmids/genetics/*metabolism ; Quinolones/*pharmacology ; beta-Lactamases/biosynthesis/genetics

BACKGROUND: There is no guideline for the appropriate wavelength at which to measure the optical density (OD) value in broth microdilution antifungal susceptibility testing, although a spectrophotometric reading method is commonly used. The present study aimed to analyze the difference in the OD values over the range of visible light and to ascertain the optimal wavelength for the spectrophotometric method of microdilution testing. METHODS: We measured the OD of background blank controls of broth medium, antifungal agents, and inocula of five type strains using a Synergy HT multi-detection microplate reader at 5-nm intervals from 380 nm to 760 nm. We also estimated the OD differences between the 50% of growth control and blank control. RESULTS: The OD of the blank control showed a parabola shape with two peaks and steadily decreased at longer wavelengths. The curves of the antifungal agent were similar to those of blank controls, and the influence of each antifungal agent on the OD was minimal. For the difference in OD between 50% of growth control and the blank control, the curve was the opposite of the blank control, and the OD increased steadily at the wavelengths above 600 nm. CONCLUSIONS: The range between 600 nm and 700 nm was the optimal wavelength for broth microdilution antifungal susceptibility testing, although any wavelength within the visible light spectrum can be used.
Antifungal Agents/*chemistry ; Azoles/*chemistry ; Culture Media/*chemistry ; Flucytosine/*chemistry ; Microbial Sensitivity Tests ; Spectrophotometry/*methods

PURPOSE: In managing acutely dyspneic patients, differentiating the underlying disease rapidly is important but not easy. Although B-type natriuretic peptide (BNP) is generally accepted as a useful marker, inconclusive results require an emergency physician (EP) to have something more confirmative. We evaluate whether Tissue Doppler Echocardiography (TDE) performed by an EP can better discriminate between heart disease and lung disease than can BNP in the emergency department (ED). METHODS: For enrolled ED patients with acute dyspnea and unclear pathology, initial BNP level and TDE performed by EP were checked prospectively. The ratios of peak early diastolic transmitral blood flow velocity (E) versus the peak early diastolic tissue velocity over mitral annulus (Ea) on TDE were recorded. The sensitivity and specificity of tissue Doppler parameters and BNP levels for diagnosing acute heart failure were calculated and we compared the discriminatory ability of the two tools. RESULTS:49 patients (39 heart failure, 10 respiratory disease) were enrolled. The area under the ROC curves for BNP and E/Ea were 0.946 and 0.888 (p<0.001) respectively. Cutoff values were 350 pg/ml for BNP (sensitivity and specificity of 82.1% and 100%) and 9.0 for E/Ea (89.2% and 100%). Especially in the group with low BNP (<350), BNP was a poor discriminator of the underlying disease, whereas E/Ea was still effective (AUC: 0.943, p=0.021). CONCLUSION: TDE by EP is a useful tool for diagnosing acute heart failure in ED and could easily and rapidly discriminate the underlying disease of acutely dyspneic patients, especially in patients with inconclusive BNP levels.
Blood Flow Velocity ; Diagnosis, Differential ; Dichlorodiphenyldichloroethane ; Dyspnea ; Echocardiography ; Echocardiography, Doppler ; Emergencies ; Heart Diseases ; Heart Failure ; Humans ; Lung Diseases ; Natriuretic Peptide, Brain ; Prospective Studies ; ROC Curve ; Sensitivity and Specificity

PURPOSE: Ultrasound is of proven accuracy in abdominal and thoracic trauma and may be useful for diagnosing extremity injury in situations where radiography is not available, such as disasters and military and space applications. However, the diagnosis of fractures is suggested by history and physical examination and is typically confirmed with radiography. As a alternative to radiography, we prospectively evaluated the utility of extremity ultrasound performed by trained residents of emergency medicine (EM) one patient with wrist and ankle extremity injuries. METHODS: Initially, residents of EM performed physical examinations for fractures. The emergency ultrasound (EM US) was performed by trained residents, who used a portable ultrasound device with a 10- to 5-MHz linear transducer, on suspected patients before radiography examination. The results of emergency ultrasound and radiography and the final diagnosis were recorded, and correlations among them were determined by using Kappa`s test. RESULTS: Thirty-nine patients were enrolled in our study. The average age was 36.6+/-19.3 years. There were radius Fx. (n=21), radius-ulna Fx. (n=1), ulna Fx. (n=1), and contusion (n=2) injuries among the wrist injury and lat.-med. malleolar Fx. (n=13), lat. malleolar Fx. (n=6), and med. malleolar Fx. (n=3) injuries among the ankle injury. Comparing EM US with radiography, we found the sensitivity, specificity, positive predictive value, and negative predictive value of EM US for Fx. diagnosis to be 100%, 66.7%, 97.3%, 100% and those of radiography to be 97.2%, 100%, 100%, and 75%, respectively. Kappa`s test for a correlation between the Fx. diagnosis of EM US and the final diagnosis of Fx was performed, and Kappa`s value was 0.787 (P = 0.004). CONCLUSION: EM US for Fx. can be performed quickly and accurately by EM residents with excellent accuracy in remote locations such as disaster areas and in military and aerospace applications. EM US was as useful as radiography in our study and had a high correlation to the final diagnosis of Fx. Therefore, ultrasound should performed on patients with extremity injury to determine whether extremity evaluation should be added to the FAST (focused abdominal sonography trauma) examination.
Ankle Injuries* ; Ankle* ; Contusions ; Diagnosis ; Disasters ; Emergencies* ; Emergency Medicine ; Extremities ; Humans ; Military Personnel ; Physical Examination ; Prospective Studies ; Radiography ; Radius ; Sensitivity and Specificity ; Transducers ; Ulna ; Ultrasonography* ; Wrist Injuries ; Wrist*

A complete cDNA sequence encoding a pore-forming subunit (Kir6.2) of ATP-senstive potassium channel in the adult worm, Clonorchis sinensis, termed CsKir6.2, was isolated from an adult cDNA library. The cDNA contained a single open-reading frame of 333 amino acids, which has a structural motif (a GFG-motif) of the putative pore-forming loop of the Kir6.2. Peculiarly, the CsKir6.2 shows a lack-sequence structure, which deleted 57 amino acids were deleted from its N-terminus. The predicted amino acid sequence revealed a highly conserved sequence as other known other Kir6.2 subunits. The mRNA was weekly expressed in the adult worm.
Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Clonorchis sinensis/*genetics/metabolism ; Helminth Proteins/*genetics/metabolism ; Human ; Molecular Sequence Data ; Potassium Channels, Inwardly Rectifying/*genetics/metabolism ; RNA, Helminth/chemistry/genetics ; Sequence Alignment ; Support, Non-U.S. Gov't

BACKGROUND: A peripheral blood smear has been the gold standard method for the diagnosis of malaria infection. Recently, many other methods have been introduced, although having inferior sensitivity and specificity to peripheral blood smears. We evaluated Neodin malaria PCR kit and its applicability in clinical settings. METHODS: Samples from seventy patients who visited Korea University hospital were used for evaluation. DNA from EDTA blood was tested in nested multiplex PCR and 470 bp for Plasmodium vivax or 340 bp for Plasmodium falciparum was confirmed after electrophoresis. The detection limit was determined by dilution of malaria positive blood with normal blood. RESULTS: Thirty-five cases of P. vivax and 10 cases of P. falciparum were noted. Except for a case of falciparum malaria, all positive cases were consistent with the peripheral blood smear results. Detection limit was 3.6 parasite/microL. CONCLUSIONS: Neodin malaria nested multiplex PCR has high sensitivity and the ability for species discrimination and may be available in the diagnosis of malaria infection.
Diagnosis* ; Discrimination (Psychology) ; DNA ; Edetic Acid ; Electrophoresis ; Humans ; Korea ; Limit of Detection ; Malaria* ; Multiplex Polymerase Chain Reaction* ; Plasmodium falciparum ; Plasmodium vivax ; Polymerase Chain Reaction ; Sensitivity and Specificity

Acute compartment syndrome is usually caused by local vascular and traumatic involvement. Thyroid disease is not a common cause of compartment syndrome and only two cases of spontaneous compartment syndrome associated with hypothyroid myopathy have been reported. Diabetic muscular infarction is a rare microvascular complication usually occured in type 1 diabetes patients under pjoor diabetic control and intramuscular volume expansion caused by muscle necrosis and increased tissue fluid can iduce compartment syndrome. We report a case of atypical bilateral anterior tibial compartment syndrome and foot drop in a 51-year-old patient with type 2 diabetes in hypothyroid state.
Compartment Syndromes* ; Diabetes Mellitus ; Foot ; Humans ; Hypothyroidism ; Infarction* ; Middle Aged ; Muscular Diseases ; Necrosis ; Thyroid Diseases

No abstract available.
Animals ; Babesiosis* ; Clindamycin ; Humans ; Male* ; Quinine

No abstract available.
Animals ; Babesiosis* ; Clindamycin ; Humans ; Male* ; Quinine