BACKGROUND: Mycobacterial culture is a confirmatory test to detect M.tuberculosis, but it takes at least 6 weeks to diagnose. PCR is a rapid and sensitive method, but it is known that PCR has a high false positive rate due to contamination, and a high false negative rate due to inhibitors. It is also known that LCR and PCR-Hybridization, recently developed methods, are more specific methods than PCR in terms of detection M.tuberculosis. In this study, we estimated the clinical utility of in house PCR, LCR and PCR-Hybridization for the detection of M.tuberculosis. METHODS: We evaluated 75 specimens, upon which M.tuberculosis culture based testing was requested, by PCR LCR, and PCR-Hybridization and compared results. Mycobacterial culture was performed on 3% Ogawa media for 8 weeks, and an in house PCR, LCx Mycobacterium tuberculosis assay kit(Abbott Laboratories, North Chicago, III) and the AMPLICOR M.tuberculosis test kit(Roche Molecular Systems, Inc. Branchburg, NJ, USA). RESULTS: In the view of the culture results, the sensitivities of the three tests were 40%, 80%, and 100% and their specificities were 98.6%, 94.3%, and 94.3%. CONCLUSION: LCR and PCR-Hybridization and rapid and sensitive methods for detecting M.tuberculosis in clinical laboratories.
Mycobacterium tuberculosis ; Polymerase Chain Reaction* ; Tuberculosis*

Cytomegalovirus (CMV) reactivation in immune compromised patients such as those undergoing hematopoietic progenitor cell transplantation (HPCT) and those with HIV infections can cause severe morbidity and mortality despite treatment with appropriate antiviral agents. The recovery of Cytomegalovirus (CMV) specific cytotoxic T lymphocytes (CTL) plays an important role in the reconstitution of CMV specific immunity in immunocompromised patients. Recent studies have reported that CMV reactivation can be successfully treated by adoptive transfer of CMV-specific T cell clones from CMV seropositive donors expanded in vitro with CMV infected fibroblasts or lysates of CMV infected cells. Other studies have used immune dominant CMV proteins or peptides to expand CMV-specific cytotoxic T lymphocytes. This review describes the clinical manifestations of CMV disease in immunocompromised patients, recent advances of antiviral therapy for CMV disease, the principals of the induction of cellular immune response to CMV, and the clinical application of CMV immunotherapy.
Cytomegalovirus Infections/*immunology/*therapy ; Humans ; *Immunocompromised Host ; *Immunotherapy, Adoptive

No abstract available.
Aged, 80 and over ; Ascorbate Oxidase/*metabolism ; Ascorbic Acid/administration & dosage/blood/*chemistry ; Breast Neoplasms/pathology ; Cholesterol/*blood ; *Colorimetry ; Enzyme Assays ; Female ; Humans ; Injections, Intravenous ; Intestine, Small/surgery ; Kidney/physiopathology ; Male ; Middle Aged ; Palliative Care ; Recurrence

BACKGROUND: We evaluated the analytical performance of the Abbott i-STAT CHEM8+, a point-of-care testing system that measures 8 basic chemical analytes, namely, sodium, potassium, chloride, total carbon dioxide, BUN, creatinine, glucose, and ionized calcium. METHODS: The precision and linearity of 8 analytes were evaluated according to the CLSI guidelines EP15-A and EP6-A, respectively, using standard material provided by the manufacturer. i-STAT CHEM8+ and other primary methods (e.g. Hitachi Clinical Analyzer 7600 for 7 analytes, Nova CCX for ionized calcium) were also compared according to the CLSI guideline EP9-A2, using 113 patient samples. RESULTS: The standard deviation (SD) of within-run and total precision of 7 analytes except chloride was within the claimed SD or within the verification value. The coefficient of variation of total precision of 7 analytes except creatinine was within 2%. With regard to linearity, all 8 analytes showed first-order equation or at least no statistical difference with the first-order equation. We observed that the efficiency of i-STAT CHEM8+ was comparable to that of primary methods, and that this method has potential applications in the clinical laboratory. CONCLUSIONS: i-STAT CHEM8+ showed good precision and linearity, and an efficiency comparable to that shown by routine chemistry analyzers; thus, it has potential applications in the clinical laboratory. It can provide much faster results and relatively accurate value to clinicians in need of immediate results, such as in an emergency unit or in the intensive care unit.
Calcium ; Carbon Dioxide ; Chemistry* ; Clinical Chemistry Tests ; Creatinine ; Emergency Service, Hospital ; Glucose ; Humans ; Intensive Care Units ; Point-of-Care Systems ; Potassium ; Sodium

BACKGROUND: We evaluated the analytical performance of the Abbott i-STAT CHEM8+, a point-of-care testing system that measures 8 basic chemical analytes, namely, sodium, potassium, chloride, total carbon dioxide, BUN, creatinine, glucose, and ionized calcium. METHODS: The precision and linearity of 8 analytes were evaluated according to the CLSI guidelines EP15-A and EP6-A, respectively, using standard material provided by the manufacturer. i-STAT CHEM8+ and other primary methods (e.g. Hitachi Clinical Analyzer 7600 for 7 analytes, Nova CCX for ionized calcium) were also compared according to the CLSI guideline EP9-A2, using 113 patient samples. RESULTS: The standard deviation (SD) of within-run and total precision of 7 analytes except chloride was within the claimed SD or within the verification value. The coefficient of variation of total precision of 7 analytes except creatinine was within 2%. With regard to linearity, all 8 analytes showed first-order equation or at least no statistical difference with the first-order equation. We observed that the efficiency of i-STAT CHEM8+ was comparable to that of primary methods, and that this method has potential applications in the clinical laboratory. CONCLUSIONS: i-STAT CHEM8+ showed good precision and linearity, and an efficiency comparable to that shown by routine chemistry analyzers; thus, it has potential applications in the clinical laboratory. It can provide much faster results and relatively accurate value to clinicians in need of immediate results, such as in an emergency unit or in the intensive care unit.
Calcium ; Carbon Dioxide ; Chemistry* ; Clinical Chemistry Tests ; Creatinine ; Emergency Service, Hospital ; Glucose ; Humans ; Intensive Care Units ; Point-of-Care Systems ; Potassium ; Sodium

PURPOSE: To identify new immunogenic HLA-A*33;03-restricted epitopes from the human papillomavirus (HPV) 16 E7 protein for immunotherapy against cervical cancer. MATERIALS AND METHODS: We synthesized fourteen overlapping 15-amino acid peptides and measured intracellular interferon-γ (IFN-γ) production in PBMC and CD8+ cytotoxic T lymphocytes (CTLs) after sensitization with these peptides using flow cytometry and ELISpot assay. The immunogenicity of epitopes was verified using a ⁵¹Cr release assay with SNU1299 cells. RESULTS: Among the fourteen 15-amino acid peptides, E7₄₉₋₆₃ (RAHYNIVTFCCKCDS) demonstrated the highest IFN-γ production from peripheral blood mononuclear cells (PBMCs), and CD8+ CTLs sensitized with E7₄₉₋₆₃ showed higher cytotoxic effect against SNU1299 cells than did CD8+ CTLs sensitized with other peptides or a negative control group. Thirteen 9- or 10-amino acid overlapping peptides spanning E7₄₉₋₆₃, E7₅₀₋₅₉ (AHYNIVTFCC), and E7₅₂₋₆₁ (YNIVTFCCKC) induced significantly higher IFN-γ production and cytotoxic effects against SNU1299 cells than the other peptides and negative controls, and the cytotoxicity of E7₅₀₋₅₉- and E7₅₂₋₆₁-sensitized PBMCs was induced via the cytolytic effect of CD8+ CTLs. CONCLUSION: We identified E7₅₀₋₅₉ and E7₅₂₋₆₁ as novel HPV 16 E7 epitopes for HLA-A*33;03. CD8+ CTL sensitized with these peptides result in an antitumor effect against cervical cancer cells. These epitopes could be useful for immune monitoring and immunotherapy for cervical cancer and HPV 16-related diseases including anal cancer and oropharyngeal cancer.
Amino Acid Sequence ; CD8-Positive T-Lymphocytes/immunology/metabolism ; Epitopes/*immunology/therapeutic use ; Female ; *HLA-A Antigens ; Human papillomavirus 16/*immunology ; Humans ; *Immunotherapy ; Interferon-gamma/analysis/*biosynthesis ; Leukocytes, Mononuclear/immunology/metabolism ; T-Lymphocytes, Cytotoxic/immunology/metabolism ; Uterine Cervical Neoplasms/*therapy

BACKGROUND: To collect high concentration of granulocytes for transfusion to neutropenic cancer patients with infections, we investigated the effect of G-CSF or dexamethasone as granulocyte mobilizers and 10% pentastarch (PS) as the sedimentation agent in granulocyte collection by leukapheresis. Subsequently, the therapeutic effect of the granulocyte transfusions was assessed. METHODS: Forty five leukapheresis were performed with CS-3000Plus (Baxter, Deerfield, IL, USA) using 10% pentastarch. The donors were classified into three groups according to their premedication drugs and the interface detector offset; group 1 used dexamethasone with offset 15 (n=16), group 2 used dexamethasone with offset 33 (n=16), and group 3 used G-CSF with offset 33 (n=10). We compared total collected granulocyte counts and granulocyte collection efficiency (GCE). RESULTS: The mean counts of total granulocytes collected and GCE were as follows; 0.9 0.5 x 1010 and 31.6 14.3% in group 1, 1.3 0.6 x 1010 and 39.0 14.2% in group 2, and 1.6 0.9 x 1010 and 63.9 32.2% in group 3, respectively. The counts of granulocytes collected in group 3 was significantly higher than that in group 1 (P<0.05). The GCE of group 3 was significantly higher than that of group 1 and group 2 (P<0.05). Sixteen granulocyte transfusions were performed to 11 patients. We observed successful therapeutic effects in 10 out of 16 transfusions (63%). CONCLUSIONS: G-CSF indicates greater potency than dexamethasone although its high cost is limitation of routine use as mobilizing agents and PS was an excellent red cell sedimenting agent in granulocyte collection. Large volume granulocyte transfusions allow high therapeutic effects in neutropenic patients with marrows of sufficient regenerating capacity.
Bone Marrow ; Dexamethasone ; Granulocyte Colony-Stimulating Factor ; Granulocytes* ; Humans ; Hydroxyethyl Starch Derivatives* ; Leukapheresis* ; Neutropenia ; Premedication ; Tissue Donors

BACKGROUND: Splenectomy is often performed for the patients with refractory chronic immune thrombocytopenic purpura (ITP). Still, there are no generally accepted guidelines for the minimum level of platelet count and the average requirement of platelet transfusion so that the patients can safely undergo splenectomy. We evaluated the changes of platelet count and transfusion requirements around the splenectomy in patients with chronic ITP. METHODS: We reviewed the medical records of 25 patients with chronic ITP. We compared the platelet counts at admission, immediately pre-op and several post-op days. We also investigated the number of platelet concentrates transfused around splenectomy. We determined the effect of splenectomy according to Difino's classification. RESULTS: The median platelet counts of the patients was 18x109/L (7-238x109/L) on admission and recovered to 108x109/L (22-460x109/L) on preoperation day by platelet transfusion and immunosuppressive treatment. The platelet counts were rapidly recovered after splenectomy from the day of operation. Only 3 patients needed platelet transfusion after splenectomy. Thirteen among twenty five patients (52%) underwent operation without platelet transfusion support. Most transfusions were done before the surgery and 80% (12/15) of the patients required transfusion of more than 10 units of random donor platelet concentrate. Twenty one patients (84%) showed the complete remission after splenectomy. CONCLUSION: Splenectomy can lead to rapid remission even in most cases of refractory chronic ITP. Many cases can undergo the operation only with treatment modalities other than transfusion such as immunosuppressive agents and/or immunoglobulin. The minimum level of platelet counts for splenectomy may be safe over 50x109/L and about 10 units of platelet concentrate may be enough for preparation of splenectomy.
Blood Platelets* ; Classification ; Humans ; Immunoglobulins ; Immunosuppressive Agents ; Medical Records ; Platelet Count ; Platelet Transfusion* ; Purpura, Thrombocytopenic, Idiopathic* ; Splenectomy* ; Tissue Donors

BACKGROUND: Among the many methods estimating the quantity of beta-hCG for pregnancy testing in urine, immunochromatography is one of most widely used semi-quantitative detection method for its convenience to use and also for its rapid result reporting system. PREG-Q(TM) is a newly introduced semi-quantitative immunochromatography method for detecting b-hCG. Clinical usefulness of PREG-Q(TM) was evaluated as a screening test for early pregnancy detection. METHODS: Accuracy, detection limit, cross-reactivity with various glycoprotein hormones, interference study, and comparison study using total 100 urine samples from pregnant (50 samples) and non-pregnant women (50 samples) was evaluated. RESULTS: All the 50 urine samples of pregnant women showed positive results, and another 50 urine samples of non-pregnant women showed negative results with PREG-Q(TM). The lower detection limit of PREG-Q(TM) was 25 mIU/mL and the result was not affected by addition of glycoprotein hormones tested. Interfering substance causing false negative or false positive results enrolled didn't affect the test results in this study. CONCLUSIONS: We conclude PREG-Q(TM) is an excellent test kit for pregnancy test, and is valuable especially for detecting early pregnancy.
Female ; Glycoproteins ; Humans ; Immunochromatography ; Limit of Detection ; Mass Screening* ; Pregnancy Tests ; Pregnancy* ; Pregnant Women

BACKGROUND: Heat shock proteins (HSPs), or stress proteins, are immunodominant antigens of many microorganisms. In this study, we have detected the anti-HSP 70 antibody and tried to explain the role of the antibody with respect to the pathogenesis of SLE. Furthermore, we have attempted to find out the possibility to link the presence of the autoantibody with the monitoring and diagnosis of systemic lupus erythematosus (SLE). METHODS: A total of 80 samples from 55 SLE patients were screened for the presence of anti-HSP 70 antibodies. Simultaneously 59 healthy people were tested as a control group. The anti-HSP 70 antibodies were measured by enzyme-linked immunosorbent assay (ELISA) and confirmed by western blot in anti-HSP 70 antibody ELISA positive samples. The activity of disease state was confirmed by the patients' medical record and systemic lupus activity measure (SLAM). RESULTS: The mean optical density (O.D.450) of ELISA in healthy controls and SLE patients were 0.15+/-0.18 (mean+/-S.D.) and 0.13+/-0.14. The correlation of SLAM Score and ELISA O.D. was r2=0.19, P=0.014. And, the mean O.D. value of ELISA was 0.18+/-0.02 and 0.11+/-0.01 before and after treatment (P <0.05). We compared samples with SLAM Score. The O.D. of anti-HSP 70 ELISA in these patients were 0.20+/-0.02 and 0.08+/-0.002 before and after treatment respectively (n=10, mean+/-S.D., P <0.01). CONCLUSIONS: Anti-HSP 70 antibody was not a clinically useful diagnostic marker in SLE patients. However, the titer of anti-HSP 70 antibody can be used for the monitoring of the therapeutic effectiveness in these patients.
Antibodies ; Blotting, Western ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Heat-Shock Proteins ; Humans ; Immunodominant Epitopes ; Lupus Erythematosus, Systemic* ; Medical Records