Twenty one units of platelet-rich plasma(PRP) were prepared from healthy volunteer blood donors, and each unit of the PRP was divided into two aliquots by using transfer bags. Using SEPACELLTM leukocyte-removal filters, each one aliquot of the PRP was filtered immediately after preparation, and the other aliquot was filtered after a 48-hour storage at a room temperature with continuous agitation. Belbre and after filtration, platelet numbers and two platelet activation markers, CD62 and CD63, were measured using hematologic autoanalyzer and'flow cytometry, respectively. The results were as follows: 1. The platelet numbers in the PRP were reduced after filtration. 2. On the point of the preparation of PRP, the mean percentage of CD63-posititve platelets(s32.86+/- 11.3.5%) was highehr than that of CD62-positive platelets(14.63+/-11.22%). 3. When filtered immediately after preparation of PRP, the CD62-positive platelets were significantly reduced(13.23+/-10.43%), however, CD63-positive platelets were not significantly reduced(29.83+/-11.05%). 4. After 48-hour storage, both two activation markers were increased, and the markers were significantly higher in the PRPs stored after filtration than in those stored without filtration. In conclusion CD63 would be a more sensitive platelet activation marker than CD62, and the platelets expressing CD62 seemed to be removed more than those expressing CD63 during filtration.
Blood Donors ; Blood Platelets* ; Dihydroergotamine ; Filtration ; Healthy Volunteers ; Humans ; Platelet Activation* ; Platelet Count

BACKGROUND: Eosinophil cationic protein (ECP), one of the eosinophil granule proteins released during allergic reactions, may play a major role in the allergic inflammatory process. The measurement of ECP in serum may be a useful indicator of eosinophil activity in ongoing inflammatory processes. We investigated the clinical utility of ECP measurement in serum in patients with bronchial asthma, allergic rhinitis and atopic dermatitis, after standardizing sample processing. METHODS: We measured the serum ECP levels in patients with bronchial asthma (n=38), chronic obstructive pulmonary diseases (COPD) (n=13), respiratory symptoms (n=19), allergic rhinitis (n=26), non-allergic rhinitis (n=24), and atopic dermatitis (n=10) and in normal healthy controls (n=16) by the fluoroenzyme immunoassay using Pharmacia CAP System, and evaluated the correlation between ECP level and blood eosinophil number, or ECP and IgE levels. Blood eosinophil number was counted by the automated cell counter. RESULTS: Serum ECP levels were significantly higher in patients with bronchial asthma (15.6+/- 12.6 g/L), COPD (13.3+/-7.2 g/L), allergic rhinitis (23.8+/-13.2 g/L), and atopic dermatitis (20.6+/- 18.4 g/L) than in normal controls (7.5+/-4.2 g/L) (P <0.05). ECP levels were also significantly higher in patients with bronchial asthma and COPD than in patients with simple respiratory symptoms (6.9+/-4.7 g/L), whose ECP levels did not statistically differ from those in normal controls. ECP levels were also significantly higher in patients with allergic rhinitis than in patients with non-allergic rhinitis (9.5+/-5.1 g/L), whose ECP levels did not statistically differ from those in normal controls. Serum ECP level and eosinophil number in peripheral blood were correlated only in patients with bronchial asthma (r=0.53, P <0.01) and no correlation between ECP and IgE levels was found in all of the patients. CONCLUSIONS: ECP is the one of the secretory components released from the eosinophil granule and measurement of ECP in serum might be one of the noninvasive tool to assess the activity in relation to eosinophil involvement in various allergic diseases.
Asthma ; Cell Count ; Dermatitis, Atopic ; Eosinophil Cationic Protein* ; Eosinophil Granule Proteins ; Eosinophils ; Humans ; Hypersensitivity ; Immunoassay ; Immunoglobulin E ; Lung Diseases, Obstructive ; Pulmonary Disease, Chronic Obstructive ; Rhinitis

BACKGROUND: Because laboratory tests are important elements of medical practice and many new tests using new technologies are being developed, the effective use of laboratory tests and services is clearly the joint responsibility of clinicians and clinical laboratory. Clinical laboratory is needed to adopt more active approach to advocating test strategy and interpreting results. We evaluated the reflex testing algorithms, consequently ordering the secondary tests by clinical pathologists. METHODS: We decided 36 test items, which can be odered consequently by clinical pathologists, working with communication between the laboratory and the clinic. In our laboratory, clinical pathologists consequently ordered the secondary tests after interpreting the primary screening test results, analyzed them with the same samples, and reported on the same day. We applied this consequently ordering system to the hospitalized patients. RESULTS: Consequently ordered secondary tests were 5,060 (17.8%) among 28,494 tests performed on 36 items during 6 months, which comprised 0.5% of total laboratory tests. Consequently ordering system was a successful attempt to improve the effectiveness of test utilization, in which we generated laboratory informations sequentially, reduced the turn-around time and played an active role in assuring effective utilization of the laboratory tests. CONCLUSIONS: Laboratory information including optimal test selection, development of methods to insure that the resulting data are utilized properly, and interpretation of test results, which can be provided more actively by clinical pathologists, should be used in assuring superior patient care. Consequently ordering system by clinical pathologists may be considered to be a means of providing patient services more efficiently.
Humans ; Joints ; Mass Screening ; Patient Care ; Reflex

BACKGROUND: Infection of hepatitis B virus(HBV) is one of the most important cause of the liver diseases in Korea, although HBV infection tends to be decreased. Diagnostic kits more accurately detecting HBV infection have been required'in order to diagnose and prevent the HBV infection. Recently LG Chemical Ltd. developed new diagnostic kits for HBsAg, anti-HBs and anti-HBc, using HBV from Korean patients. We evaluated these new kits by comparing them with microplate enzyme immunoassay (EIA) from BehringTU(Germany) and microparticle EIA (MEIA) from AbbottTM(USA). METHOD: Sera from 1,500 healthy blood donors and 500 patients were obtained to test HBsAg, anti- HBs and anti-HBc using diagnostic kits from AbbottTM, BehringTM and LG Chemical Ltd. We analyzed the results of 3 manufacturers and confirmed the discordant results of HBsAg by Southern hybridization after HBV PCR and those of anti-HBs by neutralization assay with HBsAg from LGTM. We also evaluated the reproducibility and detection limit. RESULTS: Of 1,500 healthy blood donors, HBsAg was positive in 34 (2.3%), representing completely the same results from 3 manufacturers. However, of 500 patients, 7 (1.4%) had discordant results; HBsAg was positive in all 7 sera tested with BehringTM and positive in only one tested with AbbottTM and LGTM, respectively. HBV DNA was not detected in all 7 discordant results of HBsAg, so false positive results totaled 7 (1.4%) with BehringTM and 1 (0.2%) with AbbottTM and LGTM, respectively. Of 2,000 sera, the results of anti-HBs and anti-HBc from 3 manufacturers were same in 1,876 (93.8%) and in 1,949 (97.5%), respectively. Results of HBsAg, anti-HBs and anti-HBc from 3 manufacturers were constant on repeating tests. When testing the detection limit, BehringTM kits for HBsAg and anti-HBs retained significantly higher sensitivity than AbbottTM and LGTM. On the other hand, BehringTM kit for anti-HBc showed significantly lower sensitivity than AbbottTM and LGTM. CONCLUSION: The diagnostic kits for HBV developed by LG Chemical Ltd. showed comparable results with those by AbbottTM or BehringTM and will be useful as screening blood donors or detecting patients with HBV infection.
Blood Donors ; DNA ; Hand ; Hepatitis B Surface Antigens ; Hepatitis B* ; Hepatitis* ; Humans ; Immunoenzyme Techniques ; Korea ; Limit of Detection ; Liver Diseases ; Mass Screening ; Polymerase Chain Reaction

No abstract available.
Blood Group Antigens*

BACKGROUND: The product of oxygen-free radicals inf1ict oxidative injuries on healthy cells. Antioxidants such as superoxide dismutase(SOD), glutathione peroxidase, and reduced glutathione(GSH) are present in almost all cells and play important roles in metabolism, transport, and cellular protection. We measured blood GSH levels in healthy controls and patients with non insulin dependent diabetes mellitus(NIDDM) for evaluation of the clinical usefulness of GSH. METHODS: Erythrocyte GSH levels were measured in fifty healthy controls and thirty NIDDM patients with diabetic retinopathies by Beutler's method. We also tested within-run precision, between-run precision, linearity and recovery rate to evaluate this method measuring erythrocyte GSH levels. RESULTS: The GSH levels (mean +/-SD) of NIDDM patients (5.03+/-0.67mumo1/Hb) were significantly lower than those of healthy control group (6.46+/-0.85mumo1/Hb)(P<0.001). The results of within-run precision and between-run precision when stored at 4degrees Cwere excellent (coefficient of variation were 2.79% and 2.42%, respectively), however, when stored at the room temperature the GSH levels were sharply declined. The linearity and recovery rate were acceptable. CONCLUSIONS: The prescision, linearity, and recovery rate of GSH measurement were excellent. The GSH levels in NIDDM patient group were reduced, and this probably contributes to the defective defense mechanism against increased oxidative stress. Additional measurement of other antioxidants such as superoxide dismutase and glutathione Peroxidase may be required to clarify the pathologic significance of glutathione metabolism in various diseases.
Antioxidants ; Diabetes Mellitus, Type 2 ; Diabetic Retinopathy ; Erythrocytes* ; Glutathione Peroxidase ; Glutathione* ; Humans ; Insulin ; Metabolism ; Oxidative Stress ; Superoxide Dismutase ; Superoxides

No abstract available.
Blood Platelets* ; Platelet Aggregation*

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a major nasocomial pathogen. The active surveillance of MRSA is essential to limit its transmission. The BD GeneOhm MRSA real-time PCR assay (Becton Dickinson Diagnostics, San Diego, USA) has been recently developed and used for same-day MRSA detection directly from nasal swab specimens. The authors of the present study compared GeneOhm MRSA PCR with culture methods to evaluate its diagnostic performance for MRSA active surveillance. METHODS: The present study was conducted on patients admitted to the ICU for six months. A total of 371 nasal swab specimens were obtained from patients at admission day and at the seven-day follow-up. The swab was streaked onto culture media, and presumptive S. aureus colonies were confirmed as MRSA using the BD Phoenix automated microbiology system (Becton Dickinson Diagnostic Systems, Sparks, USA). In addition, GeneOhm MRSA PCR was performed. For discrepant results between Gene-Ohm MRSA PCR and culture, the enrichment broth culture method was performed. RESULTS: There were 34 samples with discrepant results between the GeneOhm MRSA PCR and culture. The overall agreement was 90.7%. For the detection of MRSA, the GeneOhm MRSA PCR was 96.8% sensitive and 86.3% specific, with positive and negative predictive values of 83.9% and 97.3%, respectively. CONCLUSION: Identification of MRSA-colonized patients was achieved in as little as two hours, and the high negative predictive value of GeneOhm MRSA PCR suggests that the assay is a rapid method for the identification of persons who are not colonized with MRSA. However, due to the low positive predictive value, GeneOhm MRSA PCR combined with enrichment culture in cases of positive GeneOhm MRSA PCR is potentially useful for active MRSA surveillance activities.
Colon ; Culture Media ; Follow-Up Studies ; Humans ; Methicillin-Resistant Staphylococcus aureus ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction

BACKGROUND: The serological detection of the surface antigen (HBsAg) of hepatitis B virus (HBV) is the basis of detection of HBV infections in blood donors and patients with hepatitis. The aim of this study was to compare the performance of HBsAg enzyme-linked immunosorbent assay (ELISA) and HBsAg chemiluminescence immunoassay used in Korea. METHODS: We compared seven assays: Architect i 2000 (Abbott Laboratories, USA), Elecsys 2010 immunoanalyzer (Roche Diagnostics, Germany), Advia Centaur (Bayer Healthcare, USA), Murex HBsAg version 3 (Abbott Laboratories, USA), Enzygnost HBsAg 5.0 (DADE Behring, Germany), LG HBsAg ELISA (LG, Korea), and Genedia HBsAg ELISA 3.0 (Greencross Medical Science, Korea). We evaluated the sensitivity of each assay by testing serially diluted WHO HBsAg reference material, two seroconversion panels, and recombinant HBsAg with three mutations in the 'a' determinant. RESULTS: The lowest HBsAg level detected by each assay using WHO reference material was variable from 0.05 (Murex and Advia) to 0.2 IU/mL. When testing 21 seroconversion panels, the total number of positive samples was 15 by Murex and 14 by Architect. Murex, LG, and Architect detected all of the 3 mutant samples tested. CONCLUSIONS: Analytical sensitivity and mutant detecting ability among HBsAg commercial assays were variable and not related to the analytical methods, but related to the manufacturer's reagents. We suggest that each laboratory should select an HBsAg assay based on analytical performance, test throughput, and the applicability of full automation.
Chemiluminescent Measurements/*methods ; Enzyme-Linked Immunosorbent Assay/*methods ; Hepatitis B Surface Antigens/*blood ; Humans ; Reagent Kits, Diagnostic ; Reference Values ; Sensitivity and Specificity

BACKGROUND: Although the development of antibodies(anti-HBs) against hepatitis B surface antigen(HBsAg) generally leads to the clearance of the infecting hepatitis B virus(HBV), anti-HBs reactivity has been reported in patients with chronic hepatitis. Some reports suggested that concurrent presence of HBsAg and anti-HBs was a common serologic pattern in HBV carriers, however, the others insisted concurrence was associated with evidence of viral replication and features of active inflammation. This study was to investigate the frequency and significance of concurrent presence of HBsAg and anti-HBs among Korean HBsAg-positive patients. METHODS: HBV serological markers were analyzed by Abbott Axym. Anti-HBs titer was assessed quantitatively using 6 standard calibrators(0, 10, 50, 100, 500 & 1,000 mIU/mL), and reported as positive when more than 10 mIU/mL. RESULTS: Of 959 consecutive HBsAg-positive patients who had simultaneous anti-HBs determinations, HBsAg and anti-HBs were found concurrently in 31(3.2%). Anti-HBs was quantitated 10-100 mIU/mL in 25(80.6%) and more than 100 mIU/mL in 6(19.4%) among 31 HBsAg-positive patients with concurrent presence of anti-HBs. HBeAg positive rate of concurrent HBsAg/anti-HBs-positive patients(15/22 or 68.2%) was higher than that of HBsAg-positive patients without anti-HBs(227/562 or 40.4%). Of 31 HBsAg-positive patients with concurrent anti-HBs, 15 were patients with chronic hepatits, 5 with primary hepatocellular carcinoma and 2 with liver cirrhosis. CONCLUSIONS: Concurrent HBsAg/anti-HBs-positive patients were found in about 3% of Korean HBsAg-positive patients and their anti-HBs titers were less than 100 mIU/mL in 80% of concurrent HBsAg/anti-HBs-positive patients. In patients with chronic hepatits B who possessed HBsAg and anti-HBs above 100 mIU/mL, infections of HBV variants with mutations in the envelope gene might be suspected.
Carcinoma, Hepatocellular ; Hepatitis B ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens* ; Hepatitis, Chronic ; Humans ; Inflammation ; Liver Cirrhosis