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Mycobacterial Testing in Hospital Laboratories in Korea: Results of a Survey of 40 University or Tertiary-care Hospitals.

Mi Na KIM ; Sun Hwa LEE ; Sung Eun YANG ; Chik Hyun PAI

Korean Journal of Clinical Pathology.1999;19(1):86-91.

BACKGROUND: Although tuberculosis has often been referred to as diseases of the past, the current prevalence of active tuberculosis is still about 1% and it is 10th leading cause of death in Korea. In United States where tuberculosis has resurged since 1985, mycobacteriology laboratories have been playing a pivotal role in the control of tuberculosis by providing sensitive and rapid diagnostic tests. In Korea, the extent of mycobacteriology services at hospital laboratories has never been documented, although they too should play a central role in controlling tuberculosis. The purpose of this survey is to assess the facility and testing methods of mycobacteriology laboratories and the turnaround times of diagnostic tests. METHOD: In January 1997, the mycobacteriology laboratory of 40 tertiary or university hospitals in Korea were asked to complete a questionnaire involving mycobacterial test methods, test volume, turnaround time (TAT) for AFB stain. TATs for isolation, identification, and susceptibility testing were collected from the laboratory of Asan Medical Center. RESULTS: Of the 40 laboratories participating in this survey, AFB mciroscopy was performed at 40, cultures at 38, and susceptibility tests only at two laboratories. 38 laboratories referred susceptibility tests to other non-hospital laboratories. TATs for AFB microscopy were < or =24 hrs at 34 laboratories and 24-36 hrs in 6 laboratories. Isolation/identification and susceptibility tests for mycobacteria took 40.4+/-13.2 days and 45.7+/-12.4 days, respectively. All but one laboratory used solid media, mostly Ogawa media, for primary culture, and for AFB stain, 37 laboratories were using Ziehl-Neelsen method. For identification of AFB, only 4 laboratories were using a nucleic acid probe method, 18 laboratories biochemical tests, and 16 laboratories no identification test except AFB stain. CONCLUSION: Long TATs were the common and serious problems in mycobacteriology laboratories in Korea. There are urgent needs for optimizing their testing procedures if they were to play a role in the control of tuberculosis in this country. They include introduction of broth media for cultures and susceptibility tests as well as rapid methods for identification.
Cause of Death ; Chungcheongnam-do ; Diagnostic Tests, Routine ; Hospitals, University ; Korea* ; Laboratories, Hospital* ; Microscopy ; Prevalence ; Tuberculosis ; United States ; Surveys and Questionnaires

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E-test for Antifungal Susceptibility Testing of Candida Species.

Min KIM ; Woo Hyun LIM ; Jong Hee SHIN ; Dong Wook RYANG

Korean Journal of Clinical Pathology.1999;19(1):78-85.

BACKGROUND: Although standardized broth dilution methods for antifungal susceptibility testing are available, easier testing procedures are desirable. We evaluated the E-test (AB disk, Sweden) as a possible alternative instead of NCCLS (National Committee for Clinical Laboratory Standards) broth macrodilution method. METHODS: Fifty-two bloodstream isolates of Candida spp. (including 11 C. albicans, 13 C. tropicalis, 18 C. parapsilosis, 1 C. glabrata, 4 C. krusei, 2 C. pelliculosa, 2 C. lipolytica, and 1 C. guilliermondii) were tested. Amphotericin B and fluconazole MICs for each isolate were determined by both NCCLS broth macrodilution method and E-test. The results of E-test for Candida spp. were compared with those of NCCLS macrodilution method. For selecting plating media for E-test, we compared E-test results in two different media (RPMI and Casiton medium) using five ATCC Candida strains. RESULTS: As E-test media, we selected RPMI medium for amphotericin B and Casitone medium for fluconazole because of higher agreement with NCCLS method. The E-test and NCCLS method of 52 Candida spp. yielded a very narrow range of MICs (0.064-2.0 microgram/mL) for amphotericin B and a broad range of MICs (0.5-64 microgram/mL) for fluconazole. The agreements of E-test within one doubling dilutions of the macrodilution reference were 90.4% (24h and 48h) for amphotericin B, and 90.4% (24h) and 96.2% (48h) for fluconazole. CONCLUSION: The E-test is a valuable alternative to the NCCLS macrodilution method for amphotericin B and fluconazole susceptibility testing of Candida species.
Amphotericin B ; Candida* ; Fluconazole

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Trends of Anaerobic Bacteria Isolated from Clinical Specimens.

Hea Jung SHIN ; Myung Sook KIM ; Kyungwon LEE ; Yunsop CHONG

Korean Journal of Clinical Pathology.1999;19(1):70-77.

BACKGROUND: Anaerobic bacteria constitute a major part of the normal flora of the human skin, mucous membrane and intestinal tract, and can cause various infections. The incidence of anaerobic infections may differ greatly, depending on each country or hospital. METHODS: We evaluated the recent trends of anaerobic bacteria isolated from clinical specimens at Severance Hospital from 1986 to 1995. Specimens were cultured using thioglycollate medium and phenylethanol blood agar (PEBA) for 2-3 days under anaerobic condition. Identification of organism was based on conventional or commercial kit systems. RESULTS: During this period, a total of 2,664 isolates of anaerobic bacteria were obtained from 2,251 clinical specimens. The average number of anaerobes per specimen was 1.2. The frequent sources of isolation were specimens from the abdomen, followed by soft tissue, and head and neck. B. fragilis (46.3%) was the most frequently isolated gram-negative bacilli, and P. magnus (37.6%) and C. perfringens (18.8%) were the most frequently isolated gram-positive anaerobes. Abdominal, soft tissue, and head and neck infections were frequent clinical conditions. Among the anaerobe-positive specimens, only 16.8% yielded anaerobe alone while the remaining 83.2% revealed mixed infection with aerobic bacteria. CONCLUSIONS: It was concluded that B. fragilis is the most common species among gram-negative bacilli, and that P. magnus is the most common among gram-positive cocci. As well, the anaerobes are frequently isolated from specimens of the abdomen, head and neck, and soft tissue; and anaerobic infections are commonly mixed with aerobic bacteria.
Abdomen ; Agar ; Bacteria, Aerobic ; Bacteria, Anaerobic* ; Bacteroides fragilis ; Coinfection ; Gram-Positive Cocci ; Head ; Humans ; Incidence ; Mucous Membrane ; Neck ; Phenylethyl Alcohol ; Skin

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A Study of the mecI, mecA and femA Genes of Methicillin-Resistant Staphylococci.

Jee Young AHN ; Won Bae KIM ; Dong Wha LEE ; Kyung Won LEE ; Seok Ho CHOI ; Il Soo KIM ; Chae Hyung SEO

Korean Journal of Clinical Pathology.1999;19(1):62-69.

BACKGROUND: Staphylococcus aures and Staphylococcus epidermidis are those of the important pathogens that have revealed the increase of methicillin resistance. Because of increasing prevalence of staphylococci resistant to one or more antimicrobial agents, it is necessary to determine the antibiotic susceptibility of these microorganisms. Methicillin resistance is due to the production of PBP2', which is encoded by mecA gene in the chromosome. PBP 2' shows low affinity to the all of beta-lactam drugs. Therefore, the determination of gene is considered as a correct method for the antibiotic treatment procedures. METHOD: In order to examine effectiveness of detecting mecA and femA genes for the identification of methicillin resistant S. aureus (MRSA) and mecI gene in high-level-resistant MRSA, the presence of these genes in S. aureus and S. epidermidis was investigated by PCR. The types of genes were compared with phenotype by agar dilution method and investigated with MIC of methicillin. RESULTS: 1. The mecA gene detection was useful for the identification of MRSA with MRSA 100% and methicillin susceptable S. aureus (MSSA) 2.7% (P<0.001); and not useful for the identification of methicillin resistant S. epidermidis (MRSE) with MRSE 93.8% and methicillin susceptable S. epidermidis (MSSE) 52.9% (P>0.001). 2. The femA gene detection was useful for the S. aureus identification (P<0.001) and specificity 100% with MRSA 100% and MSSA 100%, MRSE and MSSE being 0%. 3. The mecI gene detection revealed MRSA 59.4%, and high-level-resistant MRSA was not detected in 39%. There was no validity of existence and nonexistence between the degree of methicillin MIC and the detection of mecI gene (P>0.001). CONCLUSION: The study concluded that the detection of both the mecA and femA genes from staphylococci by the PCR method had been considered as a correct and useful method in the identification of MRSA. mecI gene has been deemed as a repressor gene for high-level-resistant MRSA that is clinically useful as a standard. However, it is considered that the investigation should be done with later detected nucleotide sequencing of the mecI gene.
Agar ; Anti-Infective Agents ; Methicillin ; Methicillin Resistance* ; Methicillin-Resistant Staphylococcus aureus ; Phenotype ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity ; Staphylococcus ; Staphylococcus aureus ; Staphylococcus epidermidis

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Development of Simple Identification Method of Enterococci.

Young UH ; In Ho JANG ; Kap Jun YOON

Korean Journal of Clinical Pathology.1999;19(1):57-61.

BACKGROUND: The selection of identification (ID) system of enterococci depends mainly on the accuracy of ID system, cost of operation and convenience of testing. Commercial ID kits are easy to use but too expensive. The aim of the study was to develop a simple system for the identification of species of enterococci which are frequently isolated from clinical specimens. METHODS: Eight conventional biochemical tests selected for the simple ID method were hemolysis pattern, NaCl-esculin hydrolysis, tellurite tolerance, arginine dihydrolase, acid from arabinose, raffinose and methyl-alpha-D-glucopyranoside, and pigment production. Ninety one consecutive strains of enterococci from clinical specimens isolated during the period of April 1988 were tested by the simple ID method, and API rapid ID 32 STREP. RESULTS: Simple ID method with 15 ID codes was established to identify 13 species of enterococci. Among the 91 isolates tested, 88 (96.7%) were identified to the species level of enterococci by simple ID method. CONCLUSIONS: The simplified conventional ID method is simple, reliable and economical. Further modification may improve the accuracy of the enterococcal species identification system.
Arabinose ; Arginine ; Hemolysis ; Hydrolysis ; Raffinose

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Evaluation of Plasma and Urine Cellular Fibronectin as a Tumor Marker in Gastrointestinal Tract Cancers.

Nam Hee RYU ; Chang Ho JEON ; Hun Suk SUH

Korean Journal of Clinical Pathology.1999;19(1):52-56.

BACKGROUND: Cellular fibronectin (cFN) is derived from the cell surface and extracellular matrix. It is fragmented by proteolytic enzymes, which is released from invasive and malignant tumor cells. It is circulated in blood and body fluids, and finally excreted in urine. This study was undertaken to determine the usefulness of plasma and urine cFN as a tumor marker of gastrointestinal tract cancers. METHODS: We measured the concentration of cFN in plasma and urine samples by Fibronectin EIA kit (Takara Shuzo Co., Ltd., Shiga, Japan). Subjects were 20 healthy adults, 20 patients with benign diseases, 51 patients with stomach cancer and 22 patients with colorectal cancer. We evaluated the clinical records to compare the concentrations of cFN with stage and degree of cancer metastasis. RESULTS: At the recommend cut-off values of 21 microgram/mL in plasma and 157 ng/mgCr in urine, the sensitivity of plasma and urine cFN was 41.2% and 43.1% in gastric cancers; 40.9% and 50.0% in colorectal cancers, respectively. The specificity of urine cFN was 92.5% as compared to 67.1% for plasma cFN. The cFNs in plasma and urine were significantly elevated in cancer group and the concentrations were increased further with peritoneal seeding and distant metastasis. The sensitivity of urine cFN was 73.3% in stomach cancers and 100% in colorectal cancers with distant metastasis. CONCLUSION: The urine cFN is specific and the concentrations increase further with metastasis, and may be useful in diagnosis and monitoring of gastrointestinal tract cancers. Further detailed studies must be made in a large number of patients.
Adult ; Body Fluids ; Colorectal Neoplasms ; Diagnosis ; Extracellular Matrix ; Fibronectins* ; Gastrointestinal Neoplasms* ; Gastrointestinal Tract* ; Humans ; Neoplasm Metastasis ; Peptide Hydrolases ; Plasma* ; Sensitivity and Specificity ; Stomach Neoplasms

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Transferrin Analysis by Immunofixation for The Diagnosis of Cerebrospinal Fluid Leakage.

Moon Hee LEE ; Dong Seok JEON ; Hyo Jin CHUN ; Jae Ryong KIM

Korean Journal of Clinical Pathology.1999;19(1):46-51.

BACKGROUND: CSF can be leaked from the nose or ear due to fractures, tumors or surgical procedures in the skull base region, and the threat of impending meningitis necessitates early identification of it. Since 2-transferrin occurs practically in cerebrospinal fluid (CSF) and not in other body fluid, its detection from the rhinorrhea or otorrhea can be used for the diagnosis of CSF leakage. We carried out immunofixation-silver stain (IF-SS) method for detection of 2-transferrin in the CSF in order to know optimal identification condition of specific cerebrogenic marker. METHODS: The fresh CSF sample was collected by spinal tapping. 2-Transferrin was estimated by quantifying the total transferrin by nephelomertry (Behring, Germany). 2-Transferrin of CSF was identified by electrophoresis using Titan gel high resolution protein system (Beckman, USA), immunofixation with anti-human transferrin antibody (Dako, Denmark) and then stained with silver nitrate. Serial dilutions of CSF were performed to know the detection limit of 2-transferrin. To know the influence of blood mixing, tests for mixed specimen of serum and hemolysate in CSF were performed. To evaluate the specimen storage condition, tests for different temperature and storage time were performed . RESULTS: By IF-SS method, identification limit of 2-transferrin was 0.5 mg/dL in 1:4 diluted CSF with distilled water. And 2-transferrin could be detected in condition of mixing serum protein (7.5 g/dL) or hemoglobin (13 g/dL) with CSF up to 6 : 4. At various sample storage condition, such as 37degrees C, room temperature, and 4degrees C, band intensity decreased abruptly after 1 day, and it was not detected 5 days later. Mean while, in -20degrees C and -70degrees C, 2-transferin band was detected after 10 days. CONCLUSIONS: IF-SS method was sufficiently sensitive and specific for invalidation by blood contamination, and seems to be used as effective identification of 2-transferrin in the CSF without sample concentration, less diagnostic test for CSF leakage.
Body Fluids ; Cerebrospinal Fluid* ; Diagnosis* ; Diagnostic Tests, Routine ; Ear ; Electrophoresis ; Limit of Detection ; Meningitis ; Nose ; Saturn ; Silver Nitrate ; Skull Base ; Spinal Puncture ; Transferrin* ; Water

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The Influence of Food Ingestion and Sample Storage on Direct LDL-Cholesterol Measurement by Immunoseparation Method.

Hwan Sub LIM ; Jae Lim CHUNG ; Kwang Il PARK ; Jeong Ho KIM ; Oh Hun KWON

Korean Journal of Clinical Pathology.1999;19(1):40-45.

BACKGROUND: Elevated level of low density lipoprotein-cholesterol (LDL-C) is one of the major risk factors for the development of coronary heart disease. Direct LDL-C determination method by immunoseparation (DLDL-C) recently developed is claimed not to be influenced by food ingestion. We re-evaluated the effects of diet and storage conditions for this method. METHODS: Samples were collected from thirty-two medical college students before and after meal to study the effects of diet on this method. We compared the difference of LDL-C of filtered samples between refrigerated and frozen state. We also compared direct and indirect calculated measurements of LDL-C with ultracentrifugal beta-quantification (BQLDL-C) method. RESULTS: Morning 2-hour-postprandial specimen can be acceptable with no minimal significant bias, but afternoon 2-hour or 4-hour-postprandial specimen cannot be recommended due to significant negative bias (8.6-9.6%). Storage of filtered samples showed no significant difference between frozen and refrigerated state. Calculated LDL-C when triglyceride level is more than 400 mg/dL was not reliable due to large proportional and constant bias. In contrast, DLDL-C showed good accuracy comparing with BQLDL-C (y=0.909x+3.3, r=0.869, n=9, x=BQLDL-C, y=DLDL-C). CONCLUSION: In conclusion, morning two-hour postprandial specimens can be acceptable for DLDL-C, but afternoon postprandial specimens may not be recommended due to significant negative bias. DLDL-C seems to be reliable and useful especially for hypertriglyceridemic patients or follow-up cases of hypercholesterolemia with normal triglyceride or HDL-C levels.
Bias (Epidemiology) ; Cholesterol, LDL ; Coronary Disease ; Diet ; Eating* ; Fasting ; Humans ; Hypercholesterolemia ; Meals ; Risk Factors ; Triglycerides ; Ultracentrifugation

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Performance Evaluation of TOSOH Automated Glycohemoglobin Analyzer HLC-723GHb V A1c 2.2TM.

Gyu Young JEONG ; Man Kyoung LEE ; Jong Won KIM

Korean Journal of Clinical Pathology.1999;19(1):36-39.

BACKGROUND: We evaluated the performance of the TOSOH glycohemoglobin analyzer HLC-723GHb V A1c 2.2TM (TOSOH Corp. Kyoto, Japan), a recently introduced automated hemoglobin A1c (HbA1c) analyzer using high performance liquid chromatography (HPLC) method without sample pretreatment. METHODS: The performance characteristics evaluated were precision, linearity, comparison with VARIANTTM (Bio-Rad, Germany) and throughput following NCCLS evaluation protocols (EP5-T2, EP6-P, and EP9-T). RESULTS: The within-run and between-day CV's were 0.910 and 1.328 for low level (6.2%), 1.214 and 1.460 for middle level (8.5%), and 0.789 and 1.449 for high level (10.7%), respectively. We found the perfect linearity of HbA1c (%) from 6.5 to 10.2 (r2=0.9995). Comparison studies between A1c 2.2 and VARIANTTM yielded the following correlation equations; A1c 2.2TM = 0.9915 (VARIANTTM) + 0.1198 %HbA1c (r=0.9936, P < 0.0001). Throughput was 28.0 tests per hour for A1c 2.2TM compared with 15.2 tests for VARIANTTM, which were determined including red blood cell lysis time before sample loading for VARIANTTM. A1c 2.2TM did not need sample pretreatment. CONCLUSIONS: With the above results, A1c 2.2TM shows acceptable performance and is suitable for routine use in the clinical laboratory.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Erythrocytes

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A Case of Acute Lymphoblastic Leukemia which was Diagnosed with the Help of the Radiologic Study.

Mi Yeon CHOI ; Woong Soo LEE ; Ho Joon IM ; Hahng LEE ; Kyung Bin JOO

Korean Journal of Clinical Pathology.1999;19(1):31-35.

We report a case of acute lymphoblastic leukemia (ALL) in a 7-year-old female patient who had complained of limitation of motion and pain on knee and ankle joints 5 months ago. She had been initially diagnosed as juvenile rheumatoid arthritis (JRA) and received prednisone (PDS), which delayed the accurate diagnosis of the disease. But finally she has been diagnosed with ALL by the bone marrow aspiration and biopsy being done after the discontinuation of PDS with the help of leukemic suspicion of radiologic study.
Ankle Joint ; Arthritis, Juvenile ; Biopsy ; Bone Marrow ; Child ; Diagnosis ; Female ; Humans ; Knee ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Prednisone

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