BACKGROUND: The serum des-gamma-carboxyprothrombin (protein induced by vitamin K antagonist-II, PIVKA-II) is a useful tumor marker in addition to alpha-fetoprotein for diagnosing primary hepatocellular carcinoma (HCC). In this study, we evaluated the laboratory performance of a modified ELISA method for PIVKA-II measurement adopting an automated ELISA processor in comparison with conventional manual method and investigated its diagnostic performance in patients with HCC. METHODS: The laboratory performance of modified ELISA using PIVKA-II ELISA kit (Sanko Junyaku Co., Japan) was evaluated using control materials (10, 25, 500, 1,000 mAU/mL) and 208 patient samples according to the CLSI guidelines. In 93 HCC patients and 88 disease controls (30 chronic hepatitis and 58 liver cirrhosis), ROC curve, sensitivity, specificity, and positive and negative predictive values were analyzed. RESULTS: Total and within-run CVs for middle, high and very high level samples were less than 10%, while those of low level samples were over 10% (12.6% and 11.7%, respectively). The modified ELISA showed an excellent linearity (r>0.99) and low carryover rate (-0.14%). Although the correlation between the conventional and modified ELISAs was excellent (r=0.982), there was a proportional deviation of PIVKA-II levels (y intercept: 0.621). With a cut-off of 30 mAU/mL, the sensitivity and specificity of PIVKA-II for the diagnosis of HCC were 58% and 92%, respectively. CONCLUSIONS: PIVKA-II measurement by modified ELISA using an automated ELISA processor can improve the efficiency of laboratory in terms of turnaround time and labor intensiveness while maintaining reasonable sensitivity and specificity for the diagnosis of HCC.
alpha-Fetoproteins ; Biomarkers ; Carcinoma, Hepatocellular ; Enzyme-Linked Immunosorbent Assay ; Hepatitis, Chronic ; Humans ; Immunoassay ; Liver ; Protein Precursors ; Prothrombin ; ROC Curve ; Sensitivity and Specificity ; Vitamin K

BACKGROUND: Plasma cell neoplasm is diagnosed by performing bone marrow examination, serum- and urine-protein electrophoresis, and quantification of free light chains of immunoglobulins. We characterized and quantified monoclonal proteins typical of different diagnosed conditions to determine the best screening test(s). METHODS: We retrospectively reviewed diagnosis of and the characteristics of monoclonal proteins from 113 patients with monoclonal gammopathy. Monoclonal proteins were detected by agarose-gel electrophoresis and capillary electrophoresis, and if the results were ambiguous, they were confirmed by immunofixation electrophoresis. Free light chains were measured using nephelometry. RESULTS: The concentrations of monoclonal proteins in 113 patients with different conditions were as follows: multiple myeloma (MM) (67%), 2.66 (0.87-9.48) g/dL; monoclonal gammopathy of undetermined significance (MGUS) (26%), 0.62 (0.08-2.95) g/dL; lymphoma (3%), 3.65 (1.59-6.54) g/dL; Waldenstrom's macroglobulinemia (2%), 1.99 (1.08-2.90) g/dL; amyloidosis (2%), 0.61 g/dL; and POEMS syndrome (1%), 0.99 g/dL. There was a significant difference in the concentration and kappa/lambda ratio (which was based on the immunetype of the monoclonal proteins) of the monoclonal proteins in patients with MM and MGUS (P<0.001 and P=0.004, respectively). The diagnostic sensitivity of serum-protein electrophoresis, free-light-chain assay, and bone marrow analysis was 87.6%, 84.1%, and 84.5%, respectively. The sensitivity of a combination of 2 or 3 of these tests was higher at 100%. CONCLUSIONS: A combination of protein electrophoresis with immunotyping and serum free-light-chain assay may be the best screening method for detecting monoclonal proteins since its non-invasiveness.
Amyloidosis ; Bone Marrow ; Bone Marrow Examination ; Electrophoresis ; Electrophoresis, Capillary ; Humans ; Immunoglobulins ; Light ; Lymphoma ; Mass Screening ; Monoclonal Gammopathy of Undetermined Significance ; Multiple Myeloma ; Neoplasms, Plasma Cell ; Paraproteinemias ; Plasma ; Plasma Cells ; POEMS Syndrome ; Proteins ; Retrospective Studies ; Waldenstrom Macroglobulinemia

BACKGROUND: Hemoglobin A1c (HbA1c) is a universally used parameter for monitoring glycemic control in diabetic patients. Various methods are used for the measurement of HbA1c levels. The AutoLab HbA1c reagent was recently developed for use in immunoturbidimetric assays for measurement of HbA1c levels. We evaluated the reliability of using the AutoLab HbA1c reagent with the Hitachi Clinical Analyzer 7180, an automated chemistry analyzer, for measuring HbA1c levels. METHODS: We evaluated the precision, linearity, carryover rate, and stability of the samples analyzed using Hitachi clinical analyzer 7180 with the AutoLab HbA1c reagent and compared the results with those obtained with an HPLC method performed using the Variant II Turbo analyzer. RESULTS: The CV values for within-run imprecision at low and high levels were 0.8% and 0.6%, respectively, and the CV values for between-run imprecision at low and high levels were 1.5% and 2.9%, respectively. The linearity of the results was good in the range of 4.3-12.3%, and comparison with the results obtained by Variant II Turbo showed an excellent correlation coefficient of 0.9914. The carryover rate was 0%, and the samples refrigerated at 4degrees C for 15 days were found to be stable. CONCLUSIONS: In comparison with Variant II Turbo, Hitachi clinical analyzer 7180 with AutoLab HbA1c reagent showed good precision, linearity, and carryover rate. Hence, Hitachi clinical analyzer 7180 with AutoLab HbA1c reagent may be used for the diagnosis of diabetes and for monitoring blood glucose levels in diabetic patients.
Blood Glucose ; Chromatography, High Pressure Liquid ; Hemoglobins ; Humans ; Immunoassay

BACKGROUND: Circulating tumor cell (CTC) analysis is a promising new diagnostic field for estimating the risk for metastatic relapse and metastatic progression in patients with cancer. CONTENT: Different analytical systems for CTC isolation and detection have been developed as immunocytochemical and molecular assays, most including separation steps by size or biological characteristics, such as expression of epithelial- or cancer-specific markers. Recent technical advancements in CTC detection and characterization include methods based on multiplex reverse-transcription quantitative PCR and approaches based on imaging and microfilter and microchip devices. New areas of research are directed toward developing novel assays for CTC molecular characterization. QC is an important issue for CTC analysis, and standardization of micrometastatic cell detection and characterization methodologies is important for the incorporation of CTCs into prospective clinical trials to test their clinical utility. The molecular characterization of CTCs can provide important information on the molecular and biological nature of these cells, such as the status of hormone receptors and epidermal and other growth factor receptor family members, and indications of stem-cell characteristics. This information is important for the identification of therapeutic targets and resistance mechanisms in CTCs as well as for the stratification of patients and real-time monitoring of systemic therapies. SUMMARY: CTC analysis can be used as a liquid biopsy approach for prognostic and predictive purposes in breast and other cancers. In this review we focus on state-of-the-art technology platforms for CTC isolation, imaging, and detection; QC of CTC analysis; and ongoing challenges for the molecular characterization of CTCs.
Biopsy ; Breast ; Humans ; Neoplastic Cells, Circulating ; Polymerase Chain Reaction ; Population Characteristics ; Recurrence

Roseomonas is a genus of pink-pigmented, oxidative, gram-negative coccobacilli and rarely causes opportunistic infection. We report a case of wound infection by Roseomonas species in a 53-yr-old man with alcoholic liver cirrhosis. 16S ribosomal RNA (rRNA) gene sequencing was performed to confirm the infectious agent. The patient recovered without complication after ciprofloxacin treatment. To the best of our knowledge, this is the first case of Roseomonas infection reported in Korea.
Ciprofloxacin ; Humans ; Korea* ; Liver Cirrhosis, Alcoholic ; Methylobacteriaceae* ; Opportunistic Infections ; RNA, Ribosomal, 16S ; Wound Infection

Accurate D antigen blood typing is needed owing to the clinical importance of the Rh blood group. We describe a female infant who was suspected to suffer from Rh incompatible hemolytic disease of the newborn, and who showed a strong positive direct antiglobin test (DAT) result and false red blood cell (RBC) agglutination in D typing. Using chloroquine dissociation of IgG, we confirmed that the antibodies coating her RBCs were of anti-D type. D typing with 0.8% RBC suspensions in saline using saline gel cards showed 2+ RBC agglutinations. After increasing the incubation time of dissociation by chloroquine for up to 4 hr, the dissociated RBCs began to show agglutination in both the tube technique (2+) and the gel card technique (4+) for D typing, although the DAT rest was still positive. Therefore, in order to prevent mistyping as a false-negative D blood group, whenever the D blood typing of a patient with a strong positive DAT rest does not show RBC agglutination, retesting of the D blood typing is recommended by using saline-suspended RBCs or dissociated RBCs.
Agglutination ; Antibodies ; Blood Grouping and Crossmatching ; Chloroquine ; Erythrocytes ; Female ; Humans ; Immunoglobulin G ; Infant ; Infant, Newborn* ; Phenotype* ; Suspensions

A man aged 78 yr with no history of chemotherapy or toxic exposure presented with a history of dyspnea and intermittent red urine for 3 months and several years, respectively. Hematologic data at admission were as follows: hemoglobin, 65 g/L; white blood cell count, 4.05x109/L; platelet count, 96x109/L; and reticulocyte count, 10.9%. A peripheral blood smear revealed polychromasia, nucleated red blood cells, and neutrophils with a non-lobulated nucleus. The bone marrow was hypercellular and exhibited an increase in erythroid precursors with trilineage dysplasia and our findings were suggestive of refractory cytopenia with multilineage dysplasia (RCMD). Karyotype of bone marrow cells was as follows: 45,XY,der(9;17)(p10;q10),add(18)(q11.2)[10]/45,idem,del(3)(q21)[10]. Other laboratory findings showed decreased serum haptoglobin, increased lactate dehydrogenase, and increased indirect bilirubin levels. Moreover, results of the direct/indirect antiglobulin test (Coombs' test) and paroxysmal nocturnal hemoglobinuria analysis with CD55, CD59, fluorescent aerolysin (FLAER), and CD24 were negative. Cold agglutinin and Donath-Landsteiner antibodies were not detected. This is a case of myelodysplastic syndrome (MDS) associated with hemolytic anemia and complex chromosomal abnormalities at presentation.
Anemia, Hemolytic* ; Antibodies ; Bilirubin ; Bone Marrow ; Bone Marrow Cells ; Chromosome Aberrations ; Coombs Test ; Drug Therapy ; Dyspnea ; Erythrocytes ; Haptoglobins ; Hemoglobinuria, Paroxysmal ; Karyotype ; L-Lactate Dehydrogenase ; Leukocyte Count ; Myelodysplastic Syndromes* ; Neutrophils ; Platelet Count ; Reticulocyte Count

BACKGROUND: Recently, a new automated inoculating instrument, Previ Isola(R) (bioMerieux, France) was introduced. Although there are many evaluation reports about the inoculation of urine and body fluid samples using Previ Isola(R), no evaluation has been reported for blood samples. The objectives of this study were to evaluate this instrument for the inoculation of blood samples and to compare the microbiological results with the manual loop-to-plate method. METHODS: From March 2014 to July 2014, a total of 296 non-duplicate blood samples showing positive signals on the BacT/Alert 3D system were obtained, and both manual and automated methods were used for sample inoculation. Results of the two methods were compared according to five aspects: the culture result, number of single colonies, morphology of colonies, number of re-inoculations, and time required for inoculation. RESULTS: The sensitivity and specificity of Previ Isola(R) were 98.9% and 96.6%, respectively. The positive and negative predictive values were 99.6% and 90.3%, respectively, and the total concordance rate was 98.6%. For Previ Isola(R) and the manual methods, the number of average usable single colonies per plate was 25 and 16, the number of re-inoculations was 60 and 62, and the inoculation time for 15 blood samples was 30 min and 75 min, respectively. The morphology of colonies showed no differences between the two methods. CONCLUSIONS: The automated inoculation instrument, Previ Isola(R), showed relative good concordance with manual method, with high sensitivity and high specificity for blood sample inoculation. Previ Isola(R) may be useful for inoculating specimens including blood samples.
Automation ; Body Fluids ; Evaluation Studies as Topic ; Sensitivity and Specificity

BACKGROUND: Epstein-Barr virus (EBV) is known to be the causative agent of infectious mononucleosis and EBV-related malignancies. In this study, we compared the results of three real-time PCR kits for EBV DNA assays. METHODS: A total of 300 whole blood samples submitted for quantitative EBV PCR between January 2013 and September 2014 at Severance Hospital were included. The samples were tested by using the Artus EBV RG PCR Kit (Qiagen, Germany), AccuPower EBV Quantitative PCR Kit (Bioneer, Korea), and Real-Q EBV Kit (BioSewoom, Korea). Samples with discordant results between the three kits were confirmed by direct sequencing. RESULTS: The result concordance rate and kappa coefficient (K) were 86.3% and 0.69 for Artus-AccuPower, 93.3% and 0.85 for Artus-Real-Q, and 92.3% and 0.83 for AccuPower-Real-Q, respectively. The correlations between the three kits were found to be significant, with a correlation coefficient of r=0.854 for Artus-AccuPower, -0.802 for Artus-Real-Q, and -0.977 for AccuPower-Real-Q, respectively (P<0.0001). If the real-time PCR concordant results of 258 samples and the direct sequencing results of 42 real-time PCR discordant samples were assumed to be true, the sensitivity/specificity values were 0.921/0.976 for Artus, 0.902/0.965 for AccuPower, and 0.967/1.000 for Real-Q. CONCLUSIONS: The three real-time PCR kits showed excellent sensitivities and specificities. All these kits would be acceptable for clinical and therapeutic management of EBV. However, some discordant results between the kits indicate the need for caution in clinical diagnosis and staging. Further implementation of standardized methodology would be needed for EBV DNA assays.
Diagnosis ; DNA* ; Herpesvirus 4, Human* ; Infectious Mononucleosis ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction*

BACKGROUND: The cell cycle-dependent enzyme thymidine kinase 1 (TK1) is known to increase during cancer cell proliferation and has been reported as a prognostic marker for various hematologic malignancies and solid tumors. This study aimed to determine the reference interval in Korean healthy controls and to evaluate the usefulness of TK1 as a biomarker for aggressive clinical behavior in B-cell lymphoma patients. METHODS: We enrolled 72 previously untreated patients with B-cell lymphoma and 143 healthy controls. Serum TK1 levels were measured by chemiluminescence immunoassay (Liaison(R), DiaSorin, USA). We established the reference intervals in healthy controls. The diagnostic performance of serum TK1 was studied using receiver operating characteristic (ROC) analysis, and the correlation between the cutoff level for serum TK1 and clinical characteristics of B-cell lymphoma was evaluated. RESULTS: The reference range (95th percentile) of serum TK1 in healthy controls was 5.4-21.8 U/L. There was a clear difference in TK1 levels between patients with B-cell lymphoma and healthy controls (40.6+/-68.5 vs. 11.8+/-4.4 U/L, P<0.001). The area under the curve of serum TK1 for the diagnosis of B-cell lymphoma was 0.73 (cutoff, 15.2 U/L; sensitivity, 59.7%; specificity, 83.2%). An increased TK1 level (> or =15.2 U/L) correlated with the advanced clinical stage (P<0.001), bone marrow involvement (P=0.013), international prognostic index score (P=0.001), lactate dehydrogenase level (P=0.001), low Hb level (<12 g/dL) (P=0.028), and lymphocyte count (P=0.023). CONCLUSIONS: The serum TK1 level could serve as a useful biomarker for aggressive clinical behavior in B-cell lymphoma patients.
B-Lymphocytes* ; Bone Marrow ; Cell Proliferation ; Diagnosis ; Hematologic Neoplasms ; Humans ; Immunoassay ; L-Lactate Dehydrogenase ; Luminescence ; Lymphocyte Count ; Lymphoma, B-Cell* ; Reference Values ; ROC Curve ; Sensitivity and Specificity ; Thymidine Kinase* ; Thymidine*