BACKGROUND: It is well known that Methicillin-resistant Staphylococcus aureus (MRSA) is hardly controllable organism among pathogens of nosocomial infection. The MRSA infection control measures have been initiated at a brand new tertiary care teaching hospital which was opened in June, 1994. However, the control measures did bring out little effect. In 1997, reenforcement of all control measures were practiced in intensive care units. The measures brought out a significant improvement in reducing the incidence of MRSA infection, subsequently the same control measures were implemented through-out the entire inpatient area. METHODS: The following control measures have been reenforced since March 1997: first, application of thorough surveillance of confirmed MRSA infected patients: second, providing cohort care: third, enforcing handwashing practices after patient contact; fourth, establishing infected patients isolation zone: fifth, tagging infected patient's bed and medical record, providing disinfectant spray for washing hands, identifying and treating carriers among patient contact staffs, separate disposal of contaminated wastes, and finally repeating education of nursing staff and family members of the patients. Each month the number of incidence in MRSA nosocomial infection were followed and the leu supervisors were notified the outcome. RESULTS: The incidence of MRSA infection started to decline soon after the initiation of the control measures, from 132% in March 1997 to 5.8% in July 1997. In 1998, the infection rate maintained close to 2-3%. There had been 467 MRSA infected cases (5.7%) out of 8,253 discharges during the study period; among them 319 cases were infected once; 40 cases twice; 15 cases three times: four cases four times and 1 case seven times. The order of preference of organs infected are lungs (56.3%), wounds(11.8%), blood (7.9%), and urinary tract (1.9%). The highest incidence of this infection was found in Medicine (34.8%) and Neurosurgery (22.8%) CONCLUSION: The implementation and reenforcement of infection control measures are key to successful control of nosocomial infection, in particular, hand washing of patient contact staffs and eradication of carriers could be the most effective measures.
Cohort Studies ; Cross Infection* ; Education ; Hand ; Hand Disinfection ; Hospitals, Teaching ; Humans ; Incidence ; Infection Control ; Inpatients ; Intensive Care Units* ; Critical Care* ; Lung ; Medical Records ; Methicillin-Resistant Staphylococcus aureus* ; Neurosurgery ; Nursing Staff ; Tertiary Healthcare ; Urinary Tract

BACKGROUND: Infections caused by vancomycin-resistant enterococci (VRE) are becoming increasingly prevalent throughout the world. VRE can spread by direct patient-to-patient contact as well as on the hands of personnel and contaminated environmental surfaces. The purpose of this study was to examine the incidence of VRE among total enterococci from clinical specimen and investigate the antimicrobial characteristics and resistance genotypes of isolated VRE. METHODS: A total of 790 enterococcal isolates from patients over a period of 12 months were screened for vancomycin resistance using brain heart infusion agar plates supplemented with 6 g/mL of vancomycin. The incidence of VRE among enterococcal isolates was calculated from microbiology statistics program. Twenty three isolates of VRE were tested for minimal inhibitory concentrations (MIC) of vancomycin, penicillin, and gentamicin and resistance genotypes. RESULTS: In the first half period, the incidence of VRE was 1.9%, and in the second half, the incidence increased to 7.7%. Thirteen strains were found to be highly resistant to vancomycin, penicillin and gentamicin (MIC, >128 g/mL). According to the direct PCR analyses, the frequency of vanB, vanC1, and vanC2 types was 13, 7, and 3 strains, respectively. CONCLUSIONS: Continued vigilance, strict enforcement of infection control, and curtailment of vancomycin use seem to be our best approaches to controlling this increasingly important problem. For this purposes, accurate and timely detection of vancomycin-resistance and periodic investigation for incidence are essential.
Agar ; Brain ; Genotype* ; Gentamicins ; Hand ; Heart ; Humans ; Incidence* ; Infection Control ; Penicillins ; Polymerase Chain Reaction ; Vancomycin ; Vancomycin Resistance

The traditional light microscopy has limitations for precise growth assays of malaria parasites in culture or for assessment of new compounds for antimalarial activity; the speed and high reproducibility of flow cytometry can overcome these limitations. A flow cytometric method using PicoGreen, a DNA-binding fluorochrome, was developed with optimal precision suitable for performing growth assays of low-parasitemia field isolates. In addition, intra- and inter-person reproducibility of the flow cytometric and the microscopic method were compared in order to quantitatively demonstrate the improved precision. RNase treatment contributed to the precision of the flow cytometric measurements by enhancing the signal-to-noise ratios. Coefficients of variation of the method were smaller than 10% for 0.1% or higher parasitemia samples. The intra- and inter-person coefficients of variation of the flow cytometric method were three to six times smaller than those of the microscopic method. The flow cytometric method developed in this study yielded substantially more precise results than the microscopic method, allowing determination of parasitemia levels of 0.1% or higher, with coefficients of variation smaller than 10%. Thus, the PicoGreen method could be a reliable high sensitivity assay for analysis of low parasitemia samples and might be applied to a high throughput system testing antimalarial drug activity.
*Flow Cytometry ; Fluorescent Dyes/chemistry ; Humans ; *Microscopy ; Organic Chemicals/chemistry ; Parasitemia/*diagnosis ; Plasmodium falciparum/*isolation & purification ; Reproducibility of Results ; Ribonucleases/metabolism ; Signal-To-Noise Ratio

BACKGROUND: The National Committee for Clinical Laboratory Standards (NCCLS) recommends that the analytical variability must not exceed 25% of the biological variability in automated blood cell analysis. This study was conducted to determine whether routine automated blood cell analysis by Coulter STKS (Coulter Corp., Miami, FL, U.S.A) comforms with the NCCLS's recommendations. METHODS: Routine CBC analysis with STKS was performed on 22 healthy volunteers. The tests included calculating WBC, RBC, hemoglobin, MCV, platelet, MPV, and percentages of the granulocytes, lymphocytes, and monocytes. Blood samples were collected twice in one week interval to study the total variability. For the analytical variability, blood samples from 12 subjects were tested twice immediately after venipuncture for within-run variability, and samples from 10 subjects were tested immediately and 6 hours after venipuncture for within-day variability. The analytical variability was calculated as the sum of within-run and within-day variabilities. The biological variability was calculated by subtracting the analytical variability from total variability. The ratios of analytical and biological variabilities were calculated by dividing the analytical variability by the biological variability. RESULTS: Ratios of analytical and biological variabilities were as follows: 0.22 for WBC, 0.20 for RBC, 0.21 for hemoglobin, 0.39 for platelet, 1.98 for MPV, 0.07 for %granulocyte, 0.11 for %lymphocyte, and 1.81 for %monocyte. The ratio for MCV was not obtained because the analytical variability exceeded total variability. CONCLUSIONS: The analytical variability did not exceed 25% of the biological variability in all test categories except platelet, MPV and the percentage of monocyte. Thus, it is recommended that the analytic variability of all test categories be reduced so as to be in conformity with the NCCLS' recommendations.
Blood Cells* ; Blood Platelets ; Granulocytes ; Healthy Volunteers ; Lymphocytes ; Monocytes ; Phlebotomy

BACKGROUND: The Korean Society of Laboratory Medicine (KSLM) Laboratory Inspection and Accreditation Program (IAP) has been developed after one year of study supported by a research grant from the Ministry of Health and Welfare (MOHW) of the Republic of Korea from June 1998 to May 1999 to assess objectively the quality of laboratory work and assist the laboratories in improving the quality of their work. The IAP is based on peer review and voluntary participation. The IAP has been continuously improved since the first laboratory inspection began in May 1999 and it was soon expanded nationwide. The improvement was made by updating the inspection checklists to reflect feedback from inspection activities and holding frequent inspectors training workshops. This paper describes the progress and outcome of the IAP. METHODS: The IAP has been implemented nationwide through the following steps: 1) preliminary review of application papers including laboratory quality control policies and external proficiency survey results, as well as on-site inspection by inspectors; 2) addition of newly approved "Inpatient Interpretive Summary Report"checklist (IISR); 3) inspectors training workshop for the "IISR"checklist; 4) continuation of the IAP for all checklist areas including "IISR"; and 5) the first revision of checklists. RESULTS: One hundred nineteen laboratories were accredited during the first year of the IAP. Due to the implementation of the MOHW approved health insurance reimbursement item for laboratory physicians, the "IISR"checklist was created. The mean score of the laboratory inspection results was 92.8 and hospital laboratories showed a higher score on routine testing areas, however, commercial reference laboratories showed a better score on special testing areas. The checklists were revised according to the feedback from the first round of inspections. CONCLUSIONS: The nationwide implementation of the KSLM laboratory IAP was accomplished through this study. The IAP appears to have provided a firm basis for the improvement of quality and efficiency of clinical laboratories in the country.
Accreditation* ; Checklist ; Education ; Financing, Organized ; Insurance, Health, Reimbursement ; Korea ; Laboratories, Hospital ; Peer Review ; Quality Control ; Republic of Korea

BACKGROUND: The Korean Society of Laboratory Medicine (KSLM) Laboratory Inspection and Accreditation Program (IAP) has been developed after one year of study supported by a research grant from the Ministry of Health and Welfare (MOHW) of the Republic of Korea from June 1998 to May 1999 to assess objectively the quality of laboratory work and assist the laboratories in improving the quality of their work. The IAP is based on peer review and voluntary participation. The IAP has been continuously improved since the first laboratory inspection began in May 1999 and it was soon expanded nationwide. The improvement was made by updating the inspection checklists to reflect feedback from inspection activities and holding frequent inspectors training workshops. This paper describes the progress and outcome of the IAP. METHODS: The IAP has been implemented nationwide through the following steps: 1) preliminary review of application papers including laboratory quality control policies and external proficiency survey results, as well as on-site inspection by inspectors; 2) addition of newly approved "Inpatient Interpretive Summary Report"checklist (IISR); 3) inspectors training workshop for the "IISR"checklist; 4) continuation of the IAP for all checklist areas including "IISR"; and 5) the first revision of checklists. RESULTS: One hundred nineteen laboratories were accredited during the first year of the IAP. Due to the implementation of the MOHW approved health insurance reimbursement item for laboratory physicians, the "IISR"checklist was created. The mean score of the laboratory inspection results was 92.8 and hospital laboratories showed a higher score on routine testing areas, however, commercial reference laboratories showed a better score on special testing areas. The checklists were revised according to the feedback from the first round of inspections. CONCLUSIONS: The nationwide implementation of the KSLM laboratory IAP was accomplished through this study. The IAP appears to have provided a firm basis for the improvement of quality and efficiency of clinical laboratories in the country.
Accreditation* ; Checklist ; Education ; Financing, Organized ; Insurance, Health, Reimbursement ; Korea ; Laboratories, Hospital ; Peer Review ; Quality Control ; Republic of Korea

BACKGROUND: A policy development research project entitled "Feasibility study and development of clinical pathology laboratory inspection and accreditation system and its impact" was funded by the Ministry of Health and Welfare, Republic of Korea in 1998 to standardize and improve laboratory performances, hence to accomplish cost effectiveness of laboratory testing throughout the country. METHODS: The authors developed applicable inspection standards including 1) qualification and the role of laboratory director, 2) quality control and quality improvement, 3) facility and safety, and 4) inspection application requirements and detailed checklists for each laboratory discipline were developed accordingly. The College of American Pathologists Inspection and Accreditation Program was used as the model. Checklists for laboratory areas contain questionnaires with corresponding scores. The score is assigned from 2 to 4 according to the impact of the question to the outcome of the test. Checklists are for laboratory management (203 questions), hematology (146), routine chemistry (126), special chemistry (198), urinalysis (85), microbiology (282), immunology and serology (70), blood bank (246), HLA laboratory (117), flow cytometry (102), cytogenetics (137), molecular biology (232), and independent laboratory (542). The philosophy involved in the program was fairness, consistency, courteousness, consultation, and providing guidelines for future developments. Experts' consensus on subject matter was obtained before checklists were in use. Cut-off for accreditation was based on a score of 80%. Three dry and four wet workshops were held to produce 69 trained inspectors. While conducting wet workshops, 2 CAP accredited university hospital laboratories and 1 non-accredited university hospital laboratory as well as 1 CAP accredited large commercial laboratory were inspected by using newly developed checklists. RESULTS: All 4 laboratories were accredited with the mean score of 94%. The most common deficiencies were lack of proper documentation on quality control, outdated reagents in use, etc. CONCLUSIONS: The laboratory I and A program was successfully tested for its feasibility and we confirmed that its nationwide implementation was ready.
Accreditation* ; Allergy and Immunology ; Blood Banks ; Checklist ; Chemistry ; Consensus ; Cost-Benefit Analysis ; Cytogenetics ; Education ; Financial Management ; Flow Cytometry ; Hematology ; Indicators and Reagents ; Korea* ; Laboratories, Hospital ; Molecular Biology ; Pathology, Clinical* ; Philosophy ; Policy Making ; Quality Control ; Quality Improvement ; Republic of Korea ; Urinalysis ; Surveys and Questionnaires

BACKGROUND: Results of automated clinical chemistry tests are affected by many factors including analytical variability. In 1976, the College of American Pathologists (CAP) Conference on the analytical goals in clinical chemistry recommended that analytical variability should be less than 1/4 of the appropriate biological variability to improve distinction between normal and diseased populations. This study is conducted to evaluate whether automated clinical chemisty analyses performed in our laboratory is in conformance with the CAP's recommendation. METHODS: Routine chemistry and electrolyte tests were performed using Hitachi 747 automatic analyzer on 22 healthy volunteers. Blood samples were obtained from the volunteers' same vein twice in one week interval to study the total variability. Serum samples from 12 subjects were tested in duplicate immediately after blood collection for within-run analytical variability; and samples from another 10 subjects were repeated after 6 hours for within-day analytical variability. Within-run analytical variability plus within-day analytical variability make total analytical variability. Biological variability was defined as the difference between total variability and the analytical variability. Finally, ratios of analytical and biological variabilities were calculated. RESULTS: The ratios of analytical and biological variabilities of uric acid, glucose, and K were less than 0.25. But ratios of BUN, PO4, alkaline phosphatase, total bilirubin, AST, cholesterol, ALT, Cl, and protein exceeded 0.25. The ratios of Na, Ca, albumin, CO2, and creatinine could not be calculated. CONCLUSIONS: It is suggested that the analytical processes of the automated clinical chemistry tests be improved so as to be in conformity with the CAP's recommendation.
Alkaline Phosphatase ; Bilirubin ; Chemistry ; Chemistry, Clinical* ; Cholesterol ; Clinical Chemistry Tests* ; Creatinine ; Glucose ; Healthy Volunteers ; Uric Acid ; Veins

BACKGROUND: Standard protocols are lacking for the preparation of platelet lysates (PL) as an alternative to using fetal bovine serum as a cell culture supplement. This study aimed to establish optimum conditions for preparing PL for use in cell cultures. METHODS: Cell density in three pooled platelet concentrates (PC) were adjusted to 1x10(12)/L and 2x10(12)/L. PL was prepared from PC by 1 to 3 freeze-thaw (FT) cycles. HaCaT cells were cultured in media supplemented with 5% or 10% PL. Cell numbers were estimated using a Cell Counting Kit-8 (CCK-8; Dojindo Laboratories, Japan). Growth factors were quantified by using the Luminex 200 system (Luminex Corporation, USA). RESULTS: Cell proliferation rates in the presence of PLs were similar when prepared from PCs of both cell densities. The rates were higher in media containing 5% PL than 10% PL when prepared by two FT cycles. Concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor-AB/BB (PDGF-AB/BB), PDGF-AA, and epidermal growth factor (EGF) were significantly higher in PL prepared from PC with a cell density of 2x10(12)/L than 1x10(12)/L PC. However, only VEGF and PDGF-AA concentrations in PLs were correlated with HaCaT cell counts. CONCLUSIONS: The 5% PL from PC with a cell density of 1x10(12)/L prepared by two FT cycles treatment was the most effective condition that supported steady HaCaT cell proliferation. Our finding may be useful for preparing PL-supplemented cell culture media.
Blood Platelets/chemistry/*metabolism ; Cell Line ; Cell Proliferation/drug effects ; Culture Media/pharmacology ; Epidermal Growth Factor/chemistry/pharmacology ; Humans ; Platelet-Derived Growth Factor/chemistry/pharmacology ; Vascular Endothelial Growth Factor A/chemistry/pharmacology