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Evaluation of the Phoenix Automated Microbiology System for Detecting Extended-Spectrum beta-Lactamase in Escherichia coli, Klebsiella species and Proteus mirabilis.

Kyo Kwan LEE ; Sung Tae KIM ; Ki Suk HONG ; Hee Jin HUH ; Seok Lae CHAE

The Korean Journal of Laboratory Medicine.2008;28(3):185-190. doi:10.3343/kjlm.2008.28.3.185

BACKGROUND: The aim of this study was to compare the BD Phoenix (Beckton Dickinson Diagnostic Systems, USA) extended-spectrum beta-lactamase (ESBL) test with the Clinical and Laboratory Standards Institute (CLSI) ESBL phenotypic confirmatory test by disk diffusion (CLSI ESBL test) in Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis. METHODS: We tested 224 clinical isolates of E. coli, K. pneumoniae, K. oxytoca and P. mirabilis during May 2006 to March 2007. These isolates were examined by the Phoenix and the CLSI ESBL tests simultaneously. For the isolates showing discordant results between the two tests, boronic acid disk test was performed to differentiate AmpC beta-lactamase and ESBL. RESULTS: Among the 224 clinical isolates, 75 and 79 isolates were positive for ESBL by CLSI ESBL test and Phoenix test, respectively. Having detected 4 more isolates as ESBL-producers, Phoenix test showed a 98.2% agreement with a 100% sensitivity and 97.3% specificity compared with CLSI ESBL test. Among the four false positive isolates, three were AmpC-positive but ESBL-negative. CONCLUSIONS: The BD Phoenix ESBL test was sensitive and specific, and can be used as a rapid and reliable method to detect ESBL production in E. coli, Klebsiella species, and P. mirabilis.
Automation ; Bacterial Proteins/classification/*metabolism ; Disk Diffusion Antimicrobial Tests ; Escherichia coli/drug effects/*enzymology/isolation & purification ; Humans ; Klebsiella/*enzymology ; Klebsiella oxytoca/drug effects/enzymology/isolation & purification ; Klebsiella pneumoniae/drug effects/enzymology/isolation & purification ; *Microbial Sensitivity Tests ; Proteus mirabilis/drug effects/*enzymology/isolation & purification ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; beta-Lactamases/classification/*metabolism

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Establishment of Reference Intervals of Tumor Markers in Korean Adults.

Hee Yeon WOO ; Young Jae KIM ; Hyosoon PARK

The Korean Journal of Laboratory Medicine.2008;28(3):179-184. doi:10.3343/kjlm.2008.28.3.179

BACKGROUND: The sensitivity and specificity of tumor markers for detecting cancer could be significantly changed by the reference intervals of tumor markers. We established reference intervals of tumor markers in Korean adults and evaluated its importance, since the reference intervals recommended by the manufacturers were determined in the Caucasian population and have sometimes been adopted without verification. METHODS: We established the reference intervals of alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen (CA)125, carbohydrate antigen (CA)19-9, total prostate specific antigen (TPSA), cytokeratin fragment (Cyfra)21-1, and neuron specific enolase (NSE) according to the CLSI guideline in a maximum number of 1,364 healthy adults aged 20-60 yrs who visited a health promotion center from January to February 2007. RESULTS: Reference intervals of all tumor markers except for AFP were not in agreement with those recommended by the manufacturers. Reference intervals of CEA, TPSA, CA19-9, CA125, and Cyfra21-1 were age dependent. The mean reference values of NSE, CA125, and CEA were statistically different according to gender (11.72 vs 10.78 ng/mL), menopause status (18.89 vs 12.62 U/mL), and smoking status (2.60 vs 2.12 vs 1.80 ng/mL for smokers, past smokers, and non-smokers, respectively),respectively. CONCLUSIONS: With the verification and establishment of reference intervals of tumor markers in a Korean local population, we found the reference intervals significantly different by either age, gender, smoking or menopause status.
Adult ; Age Factors ; Carcinoembryonic Antigen/analysis ; Female ; Humans ; Korea ; Male ; Menopause ; Middle Aged ; Phosphopyruvate Hydratase/analysis ; Prostate-Specific Antigen/analysis ; Questionnaires ; Reagent Kits, Diagnostic ; Reference Values ; Sex Factors ; Smoking ; Tumor Markers, Biological/*standards ; alpha-Fetoproteins/analysis

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Two Cases of Trisomy 19 as a Sole Chromosomal Abnormality in Myeloid Disorders.

Soon IL JUNG ; Hee Soon CHO ; Chae Hoon LEE ; Kyong Dong KIM ; Jung Ok HA ; Min Kyoung KIM ; Kyung Hee LEE ; Myung Soo HYUN

The Korean Journal of Laboratory Medicine.2008;28(3):174-178. doi:10.3343/kjlm.2008.28.3.174

Trisomy 19 is frequently encountered in cases of chronic myeloid leukemia (CML) as a secondary abnormality: however, trisomy 19 rarely occurs as a sole chromosomal abnormality and, to date, it has only been reported in 48 hematopoietic malignancies, 1 case of adenocarcinoma and 1 case of astrocytic tumor. Here, we report two additional cases of trisomy 19 as a sole karyotypic aberration in myeloid malignancies. One of these cases involved a 6-month-old male who was diagnosed with acute myeloid leukemia minimally differentiated. His karyotype was 47,XY,+19[20]. He expired 5 days after diagnosis. Another case occurred in an 80-yr-old female who had refractory anemia with excess blasts. Her karyotype was 47,XX,+19[16]/46,XX[4]. Four months later, her peripheral blood smears suggested that the disease had progressed, but she refused further evaluation. Based on a review of the existing literature and the results of this report, trisomy 19 not only as a secondary abnormality but also as a sole karyotypic aberration is strongly associated with myeloid disorder; however, it is not preferentially found in specific FAB subgroups of myelodysplasic syndrome or acute myeloid leukemia.
Acute Disease ; Aged, 80 and over ; Anemia, Refractory/*diagnosis/*genetics ; *Chromosomes, Human, Pair 19 ; Female ; Humans ; Infant ; Karyotyping ; Leukemia, Myeloid/*diagnosis/*genetics ; Male ; *Trisomy

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Serum Free Light Chains for Diagnosis and Follow-up of Multiple Myeloma.

Seonkyung JUNG ; Myungshin KIM ; Jihyang LIM ; Yonggoo KIM ; Kyungja HAN ; Chang Kee MIN ; Woo Sung MIN

The Korean Journal of Laboratory Medicine.2008;28(3):169-173. doi:10.3343/kjlm.2008.28.3.169

BACKGROUND: Free light chain (FLC) is widely used to evaluate B-cell proliferative diseases. Herein, we estimated the clinical usefulness of serum FLC in multiple myeloma (MM). METHODS: Fifty-one patients were enrolled. We performed FLC analysis, protein electrophoresis (PEP), and immunofixation electrophoresis (IFE). FLC was measured using Toshiba 200 FR Neo with FREELITE(TM), and kappa/lambda (kappa/lambda) ratio was calculated. We compared these parameters in 41 patients with increased FLC before and after bortezomib treatment. Complete response (CR) was defined as the disappearance of monoclonal (M) protein in serum and/or urine as measured by IFE. Partial response (PR) was defined as > or =50% reduction of serum M protein. Early objective response (EOR) included both CR and PR. Minimal response (MR) was defined as 25-49% reduction of M protein and stable disease (SD) as <25% reduction. RESULTS: Forty-one (80.4%) of the 51 patients studied revealed increment of FLC and the five patients with no increment revealed an abnormal kappa/lambda ratio. Especially, all of the light chain myeloma and non-secretory myeloma showed increased FLC concentrations. Among the patients with EOR, 72.4% (21/29) showed a normal or subnormal FLC concentration after the first cycle of treatment. Otherwise, PEP and IFE normalized in 24.1% (7/29) and 24.1% (7/29), respectively. The ratio of decreased FLC after the first cycle of treatment was significantly different between EOR and other response groups (MR, SD) (90.6% vs 51.8%, P=0.011). CONCLUSIONS: FLC was considered as a good diagnostic method in complement with PEP and IFE in MM, especially in light chain myeloma or non-secretory myeloma. Moreover, FLC is a useful monitoring tool because it reflects therapy results more rapidly owing to a short serum half-life.
Adult ; Aged ; Boronic Acids/therapeutic use ; Female ; Humans ; Immunoelectrophoresis ; Immunoglobulin Light Chains/*blood/urine ; Male ; Middle Aged ; Multiple Myeloma/*diagnosis/therapy ; Pyrazines/therapeutic use ; Reagent Kits, Diagnostic

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Reorganization of Automated Outpatient Laboratory to improve Total.

Woochang LEE ; Won Ki MIN ; Sail CHUN ; Seongsoo JANG ; Hyun Sook CHI ; Chan Jeong PARK ; Hyosoon PARK

The Korean Journal of Laboratory Medicine.2004;24(5):334-338.

BACKGROUND: Asan Medical Center ran a fully automated outpatient laboratory to serve outpatient departments with a rapid turnaround time (TAT; one hour from reception to reporting) for frequently requested test items and thus to make `one stop service' possible. As the number of samples increased, the TAT gradually became longer and eventually showed over one hour for almost all items. METHODS: In October 1998, reorganization of the outpatient laboratory took place. Newly introduced were a priority system for samples, a plasma separate tube instead of the serum separate tube, an on-line simultaneous sample reception system at the time of sampling, and a real-time monitoring system for TAT. RESULTS: With the sample priority system, samples processed for one stop service were 61.0% (476 priority samples out of 780 total samples) for routine hematology, 59.2% (527 out of 890) for routine chemistry, 40.1% (122 out of 304) for urinalysis, 43.2% (89 out of 206) for coagulation tests, and 47.9% (75 out of 157) for diabetic tests. We monitored samples processed as `one stop service' with the real time monitoring system for TAT; among the samples processed as such, 80.1% of rou-tine hematology, 91.9% of routine chemistry, 99.5% of urinalysis, 92.6% of coagulation tests, and 97.6% of diabetic tests showed TAT less than one hour. Average TAT, from the specimen acquisition to the reporting, decreased dramatically after introduction of the system. Average TAT of routine hematology decreased to 53.9 minutes, routine chemistry to 54.6 minutes, urinalysis to 35.2 minutes, coagulation tests to 46.6 minutes, and diabetic tests to 31.9 minutes. And the patient satisfaction index for the outpatient laboratory rose 15% from 81% to 96%. CONCLUSIONS: The outpatient laboratory of AMC shortened the TAT substantially without sacrificing quality and fully met the needs of patients and clinicians after reorganization.
Chemistry ; Chungcheongnam-do ; Hematology ; Humans ; Outpatients* ; Patient Satisfaction ; Plasma ; Urinalysis

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Influence of Standard Curves on Relative Quantification using Real-time PCR.

Mi Kyung LEE ; Tae Hyoung KIM

The Korean Journal of Laboratory Medicine.2004;24(5):327-333.

BACKGROUND: Although relative quantification by real-time PCR may be easier to perform than absolute quantification, there is a risk of errors associated with standard curve construction. The aim of this study was to evaluate the effects of standard curves on relative quantification using real-time PCR in Candida albicans. METHODS: The reproducibility of real-time PCR-based standard curves for target genes and a reference gene generated from PCR amplicons (10-fold serial dilution, 10(-4) to 10(-9)) was evaluated. In addition, the effects on standard curves were evaluated by running the same cDNA samples. RESULTS: The within-run variation (CV) by crossing point (Cp) was 0.12-1.05% for ERG11 and ACT1, whereas the between-run CV was 2.07-6.84% for ERG11, CDR1, MDR1 (target gene) and ACT1 (ref-erencegene). The differences in PCR efficiency between targets and reference may be attributable to variations in relative quantification. CONCLUSIONS: To achieve reliable relative quantification of mRNA in real-time PCR, a feasible guide-line and standardization are of major importance.
Candida albicans ; DNA, Complementary ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction* ; RNA, Messenger ; Running

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The Comparison of Restriction Fragment Mass Polymorphism Method with Sequecing Method for the Detection of HBV YMDD Mutants.

Yong Hak SOHN ; Soo Jin YOO ; Jin Hyuk YANG ; Sang Hyun HWANG ; Sun Pyo HONG ; Heung Bum OH

The Korean Journal of Laboratory Medicine.2004;24(5):320-326.

BACKGROUND: YMDD motif mutants of the hepatitis B virus (HBV) emerged in some chronic hepatitis B patients after prolonged lamivudine treatment. Recently a novel genotyping assay, the restriction fragment mass polymorphism (RFMP) method, was introduced for the detection of YMDD mutations. We compared the performance of the RFMP method with that of sequencing method in chronic hepatitis B patients who had suffered the HBV DNA breakthrough after lamivudine treatment. METHODS: Enrolled in this study were 18 chronic hepatitis B patients who experienced the DNA breakthrough after a period during which HBV DNA was undetectable by Hybrid capture II HBV DNA test (Digene Inc., Gaithersburg, MD, USA). Both sequencing and RFMP methods were used to detect YMDD variants in three phases such as before treatment, before breakthrough and after breakthrough. RESULTS: YMDD mutants were detected in 13 samples (72.2%) by both methods after DNA breakthrough. Among them were two samples with a mixed HBV population detected by RFMP. Before breakthrough, the mutants were detected in three samples (16.7%) by sequencing and four (22.2%) by RFMP, showing discrepant results for two samples. The concordance rate between both methods was 92.6%. CONCLUSIONS: Both sequencing and RFMP methods were highly concordant except in a few cases, so it is suggested that both methods are appropriate in detecting YMDD mutants.
DNA ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Lamivudine

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The Cryopreservation and Thawing of Red Blood Cells Using 25% Hydroxyethyl.

Tae Hee HAN ; In Ki PAIK

The Korean Journal of Laboratory Medicine.2004;24(5):314-319.

BACKGROUND: The method using hydroxyethyl starch (HES) as a cryoprotectant is a simple, quick, and inexpensive way to make frozen red blood cells (RBCs) for an extended period. It needs neither sophisticated equipment nor skilled labor. As HES is a plasma expander, it does not have to be washed out before transfusion. But, it is not yet introduced in Korea to make frozen RBCs with HES. The aim of our study was to establish a method to cryopreserve RBCs using HES in Korea. METHODS: We rejuvenated RBCs (20 units) on the first day after expiration. They were washed three times with saline adenine glucose mannitol phosphate buffer solution (SAGMP), then their hemat-ocrits were adjusted up to 80%. The same weight of 25% HES was added to each RBC, to make a final mixture of 12.5% (wt/wt) HES-40% RBC. The blood bags were placed in a thin metal frame and frozen in liquid nitrogen. After 3 months, the blood bags were thawed in a 37 degrees Cwater bath. To evaluate the qualities of frozen-thawed RBCs, samples of HES-RBC mixtures before freezing and after thawing were tested for ATP, 2, 3-DPG, potassium, plasma hemoglobin, hemolysis %, and 30-minute saline stability (30-min SS). RESULTS: After thawing, the percentage (mean+/-SD) of RBC hemolysis was 1.6+/-0.4% (range, 1.1-2.6) and three units were over 2%; 30-min SS was 92.0+/-0.16% (88.4-93.8). Supernatant potassium and free hemoglobin were 15.6+/-3.5 mmol/L (10.9-23.0) and 215+/-62 mg/dL (139-318), respectively. The ATP and 2, 3-DPG of RBCs were 4.23+/-0.81 (3.31-6.51) micromol/g Hb and 8.46+/-1.62 (5.21-11.00) micromol/g Hb, respectively. CONCLUSIONS: Of the 20 units of RBCs tested, 17 units (85%) were within the criteria for transfusion (hemolysis<2%, 30 min SS>88%, potassium<75 mmol/L) after thawing. We could successfully freeze and thaw RBCs utilizing 25% HES as a cryoprotectant.
Adenine ; Adenosine Triphosphate ; Baths ; Cryopreservation* ; Erythrocytes* ; Freezing ; Glucose ; Hemolysis ; Korea ; Mannitol ; Nitrogen ; Plasma ; Potassium ; Starch

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Experience of Bloodless Medicine and Surgery in Soonchunhyang University.

Byung Ryul JEON ; Jeong Won SHIN ; Yujin PARK ; Rojin PARK ; Tae Youn CHOI ; Hee Bong SHIN ; You Kyung LEE

The Korean Journal of Laboratory Medicine.2004;24(5):308-313.

BACKGROUND: We established a bloodless center at Soonchunhyang University Hospital (SCH) in 1996 and have provided medical and surgical care for Jehovah's Witness patients. In this study, we evaluated their outcomes to provide the basis of bloodless medicine and surgery in Korea. METHODS: A retropective review of the medical records of 757 Jehovah's Witness patients admitted in the SCH Bloodless Center from December 1996 to July 2003 was performed. RESULTS: Among 757 patients, 19 (2.5%) expired during treatment and 4 of them died of cardiopul-monary dysfunction secondary to anemia. As alternatives to blood transfusion, 85 (11.2%) patients were treated with iron, 81 (10.7%) with erythropoietin, 49 (6.5%) with aprotinin, 31 (4.1%) with hemodilution and 28 (3.7%) with cell saver. Four hundreds fifteen (54.8%) of 757 patients underwent surgery. The most frequently involved cinical department was Obstetric/Gynecology (23.8%). The ratio of female and the percentage of cases treated with alternatives to blood transfusion were higher in surgery group than non-surgery group patients.(Chi-square test, P<0.01) CONCLUSIONS: Most Jehovah's Witness patients were treated successfully in our bloodless center with various alternatives to blood transfusion, such as erythropoietin, intraoperative autotransfusion, acute normovolemic hemodilution etc. Collaboration and good communication among surgeons, anesthesiologists, hematologists and blood bank physicians are very important to provide qualified medical or surgical treatment to the patients who have a religious objection to receiving blood or blood-related products.
Anemia ; Aprotinin ; Blood Banks ; Blood Transfusion ; Blood Transfusion, Autologous ; Cooperative Behavior ; Erythropoietin ; Female ; Hemodilution ; Humans ; Iron ; Korea ; Medical Records

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Evaluation of Spectrophotometric Broth Microdilution Methods for Determining Antifungal Susceptibilities of Aspergillus Species.

Mi Ra PARK ; Jong Hee SHIN ; Jeong Won SONG ; Young Kyu PARK ; Duck CHO ; Seung Jung KEE ; Myung Geun SHIN ; Soon Pal SUH ; Dong Wook RYANG

The Korean Journal of Laboratory Medicine.2004;24(5):301-307.

BACKGROUND: A spectrophotometric approach to minimum inhibitory concentration (MIC) determination for filamentous fungi may provide an objective and rapid MIC reading, and quantify the hyphal growth of molds. In this study, we evaluated two spectrophotometric broth microdilution methods (SBM) to determine amphotericin B and itraconazole MICs for Aspergillus species isolated from clinical specimens. METHODS: A total of 80 clinical isolates (20 A. fumigatus, 20 A. flavus, 18 A. niger, 20 A. terreus, and 2 A. nidulans) were tested for amphotericin B and itraconazole susceptibility by the broth microdilution method. The MIC endpoint was calculated by the spectrophotometer with microplate reader (SBM-Spec method) or colorimetric XTT (tetrazolium dye) method (SBM-XTT method). The results of the SBM method were compared with those of NCCLS M38-A broth microdilution method. RESULTS: The MICs of amphotericin B by the NCCLS M38-A method ranged from 0.125 to 8 g/mL, and those of itraconazole ranged from 0.25 to 2micrograms/mL. The agreement of SBM-Spec and SBM-XTT methods within one dilution of the NCCLS M38 reference were 98.8% and 96.3% for the ampho-tericin B, and 98.8% and 100% for itraconazole, respectively. The agreements between SBM-Spec and SBM-XTT methods were 97.5% for amphotericin B and 98.8% for itraconazole. CONCLUSIONS: In antifungal susceptibility testing of Aspergillus species, the SBM method includ-ing SBM-Spec and SBM-XTT methods showed high levels of agreements with the NCCLS M38-A method. The SBM methods can be useful in the clinical laboratory.
Amphotericin B ; Aspergillus* ; Fungi ; Itraconazole ; Microbial Sensitivity Tests ; Niger

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