Novel Swine-Origin Influenza A (H1N1) Viral Encephalitis

The World Health Organization declared that a new strain of novel swine-origin influenza A (H1N1) virus was responsible for the pandemic infection in June 2009. We report a case of encephalitis diagnosed as the H1N1 virus infection. We describe a 17-year-old patient who had a seizure attack, diagnosed with a H1N1 virus infection via real time reverse-transcriptase polymerase chain reaction (RT-PCR). The H1N1 virus infection can be causative of the encephalitis, as with other influenza virus infections. Careful monitoring is essential for reducing complications.

Soluble syndecan-1 at diagnosis and during follow up of multiple myeloma: a single institution study / 대한혈액학회지

BACKGROUND: Syndecan-1 is a heparan sulfate proteoglycan expressed on plasma cells, especially myeloma cells, and can exist in serum as soluble syndecan-1 after shedding from the cell surface. Soluble syndecan-1 has been suggested to promote myeloma cell growth and to be an independent prognostic factor for multiple myeloma. We aimed to evaluate the effect of soluble syndecan-1 levels at the time of diagnosis and during therapy on therapeutic response and prognosis for patients with multiple myeloma. METHODS: We analyzed soluble syndecan-1 levels in 28 patients with multiple myeloma and 50 normal controls, and compared its levels with Durie-Salmon stage and other markers of myeloma. In addition, we evaluated the therapeutic response and determined the 3-year survival rates of these patients. RESULTS: We observed that the median soluble syndecan-1 level in myeloma patients was higher than that in the normal controls (P <0.0001), and the soluble syndecan-1 levels in 21 (75%) patients were higher than the cut-off level (162 ng/mL). Soluble syndecan-1 levels correlated with disease stage, percentage of plasma cells in the bone marrow, beta2 microglobulin level, serum M-component concentration, and creatinine level. The baseline levels of soluble syndecan-1 at the time of diagnosis in the patients who responded to chemotherapy were lower than those in the non-responders (P=0.04); however, the baseline level was not a significant predictor of therapeutic response. The 3-year overall survival rate of the patients with high soluble syndecan-1 levels at the time of diagnosis and 6 months after chemotherapy was lower than the corresponding survival rates of the patients with low levels of soluble syndecan-1; however, the overall survival rate was not statistically significant. CONCLUSION: The use of soluble syndecan-1 has limitations in the diagnosis of multiple myeloma. Soluble syndecan-1 levels correlate with known prognostic factors; however, we could not assess the prognostic value of high levels of soluble syndecan-1 at the time of diagnosis and after chemotherapy.

The M142T Mutation Causes B3 Phenotype: Three Cases and an in vitro Expression Study / 대한진단검사의학회지

The B3 phenotype is the most common B subtype in Korea. The B305 allele (425 T>C, M142T) was first reported in 2 Chinese individuals; however, it has not yet been reported in the Koreans, and the impact of the M142T mutation on the expression of the B3 phenotype has also not been studied. To resolve an ABO discrepancy between a group O neonate and her group O father and A(1)B(3) mother, blood samples from these individuals and other family members were referred to our laboratory for ABO gene analysis. The B305 allele was discovered in the neonate (B305/O01), her mother (A102/ B305), and her maternal aunt (B305/O02), while her father was typed as O01/O02. Transient transfection experiments were performed in HeLa cells using the B305 allele synthesized by site-directed mutagenesis; flow cytometric analysis revealed that this transfect expressed 35.5% of the total B antigen produced by the B101 allele transfect. For comparison, Bx01 allele transfects were also created, and they expressed 11.4% of the total B antigen expressed on the surface of B101 transfects. These experiments demonstrate that the M142T (425 T>C) mutation is responsible for the B subtype phenotype produced by the B305 allele.

Annual Report on External Quality Assessment in Hematology in Korea (2008) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2008 with average 822 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 96.5%. The coefficients of variation in hemoglobin, hematocrit and RBC was stable but variable in platelet count and WBC count according to measuring cell count. Test results of blood cell morphology showed variation among various cell morphologies.

A case of primary malignant uterine cervical Non-Hodgkin's lymphoma with breast involvement / 대한내과학회지

Primary lymphoma arising from the uterine cervix has been rarely encountered, and breast involvement is rare because of the relative paucity of lymphoid tissue in the breast. A 32-year-old woman with a primary malignant lymphoma of the uterine cervix and breast involvement is reported. The patient presented with post-coital vaginal bleeding, and punch biopsy of the cervix revealed the diffuse large B cell type of malignant lymphoma. PET-scan was done for staging, and abnormal FDP uptakes were detected in a uterine cervical mass and breast nodule. Ultrasonography-guided needle biopsy was done for the breast mass, and 2 biopsied nodules revealed fibroadenoma and diffuse large B cell lymphoma. The patient (Ann Arbor stage IV) was treated with 6 cycles of combination chemotherapy with CHOP plus rituximab. The patient went into complete remission. Thereafter, 4500cGy pelvic irradiation was done for adjuvant therapy.

Annual Report on External Quality Assessment in Hematology in Korea (2007) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2007 with average 722 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 95.2%. The coefficients of variation in hemoglobin, hematocrit and RBC were stable but variable in platelet count and WBC count according to measuring cell counters. Test results of blood cell morphology showed variation among various cell morphologies.

Annual Report on External Quality Assessment in Hematology in Korea (2007) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2007 with average 722 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, hematocrit, red blood cell count, platelet count, blood cell morphology, prothrombin time and activated partial thromboplastin time. The response rate was more than 95.2%. The coefficients of variation in hemoglobin, hematocrit and RBC were stable but variable in platelet count and WBC count according to measuring cell counters. Test results of blood cell morphology showed variation among various cell morphologies.

Detection of Anti-ENA and anti-dsDNA Antibodies Using Line Immunoassay in Systemic Autoimmune Diseases / 대한진단검사의학회지

BACKGROUND: Detection of antibodies to extractable nuclear antigens (ENAs) and dsDNA is needed for the diagnosis of and predicting prognosis in systemic autoimmune diseases. Recently introduced line immunoassay (LIA) has the advantage of detecting several autoantibodies simultaneously, and we evaluated its usefulness in the diagnosis of autoimmune diseases in comparison with enzyme-linked immunosorbent assay (ELISA). METHODS: Samples were collected from 437 patients referred by rheumatologists. FANA (fluorescent antinuclear antibody) test and LIA for the detection of 13 different autoantibodies, including 6 ENAs and dsDNA were performed. LIA-positive samples for ENA or dsDNA antibodies were further tested with ELISA. Final diagnosis was made by rheumatologists according to the diagnostic criteria. Agreement of results between LIA and ELISA was analyzed in 53 selected patients with systemic autoimmune diseases. RESULTS: The LIA detected antibodies to ENA and dsDNA in 118 and 22 patients, respectively, and ELISA detected 70.3% (83/118) and 45.5% (10/22) of LIA positive samples. Especially, 60.2% (71/118) of patients with positive ENA antibody on LIA was diagnosed as systemic autoimmune diseases. Patients having strong FANA titer and homogenous/speckled pattern showed higher prevalence of autoantibodies, but a small proportion of FANA negative patients also showed positive reactivity (LIA 10.8%, ELISA 5.2%). LIA showed a good agreement with ELISA for the anti-ENA antibodies (> or =80%), and a lower agreement for the anti-dsDNA antibody (67.9%). CONCLUSIONS: LIA detecting several autoantibodies simultaneously might replace ELISA for anti-ENA antibodies, but not for anti-dsDNA antibodies. When LIA is performed considering clinical manifestations and FANA, it could contribute to the diagnosis of systemic autoimmune disease.

Evaluation of Urine NMP22 Point-of-Care Test for the Screening of Bladder Cancer / 대한진단검사의학회지

BACKGROUND: Screening of high-risk patients using bladder tumor markers can offer an advantage of early detection and saving medical costs. For these purpose many tumor markers have been developed to supplement invasive cystoscopy. Our study evaluated the NMP22 point-of-care test (NMP22 POCT), which is one of the tumor makers, comparing with the standard urine cytology for the diagnosis of bladder cancer. METHODS: From January to September 2005, 232 patients who had undergone a cystoscopy due to bladder cancer associated symptoms including hematuria and dysuria were enrolled in this study. Urine specimens were collected for NMP22 POCT and cytology. NMP22 POCT and urine cytology were compared for sensitivity and specificity. In addition, we evaluated urine stick test and microscopy to explain some false-positive results in NMP22 POCT. RESULTS: Superficial transitional cell carcinoma was diagnosed in 10 patients. The sensitivity of NMP22 test was 60% (95% confidence interval [CI], 26.2-87.8%), whereas that of cytology was 33.3% (95% CI, 7.5-70.1%); however, the difference was not significant. The specificity of NMP22 test was 69.8% (95% CI, 63.3-75.8%), compared with 99.0% (95% CI, 96.5-99.9%) for cytology (P<0.001). The presence of microscopic RBCs in urine specimen was significantly associated with the lower specificity of NMP22 POCT (P=0.02). CONCLUSIONS: NMP22 POCT was significantly less specific than urine cytology. To be useful as a bladder cancer screening test, the NMP22 test should have a higher specificity.

Annual Report on External Quality Assessment in Hematology in Korea (2005) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2005 with about 500 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, red blood cell count, platelet count, white cell differential count, red blood cell morphology. The response rate was more than 97%. The coefficients of variation in hemoglobin and RBC number was stable but variable in platelet number and WBC number according to measuring cell counts. Test results showed wide variation according to measuring machine and reagents.

Diagnostic Performance of the Antifilaggrin Antibody in Rheumatoid Arthritis / 임상검사와정도관리

BACKGROUNDS: Rheumatoid factor (RF) is common serological marker for the diagnosis of rheumatoid arthritis (RA), but its sensitivity and specificity are not satisfactory for the diagnosis of RA. Therefore, we investigated the diagnostic performance of a new antifilaggrin antibody test by enzyme linked immunosorbent assay (ELISA) in RA. METHODS: Recombinant human filaggrin was deiminated in vitro by peptidylarginine deiminase and used as the coating antigen for ELISA. We performed the RF and the antifilaggrin antibody for 324 RA patients, 251 non-RA patients (rheumatic diseases other than RA), and 286 normal individuals and evaluated the sensitivities and specificities of RF and antifilaggrin antibody. Optimal cut off values were calculated as mean+2SD in 95% confidence interval except 3SD for 286 normal individuals. Optimal cut off values of antifilaggrin antibody and RF were 9.6 U/ml and 12 U/ml, respectively. RESULTS: The sensitivities and specificities of antifilaggrin antibody were 44.8% and 89.2% at optimal cut off values. The sensitivity and specificity of RF were 75.0% and 83.3%. Combination of "antifilaggrin antibody and RF" showed significantly high specificity of 95.2% and that of "antifilaggrin antibody or RF" showed slightly high sensitivity of 79.3% at optimal cut off values. Antifilaggrin antibody was positive in 17.3% among 81 sero-negative RA patients. CONCLUSION: We considered that antifilaggrin antibody could be used a supplementary test of RF for the diagnosis of RA, because "antifilaggrin antibody and RF" had higher diagnostic specificity than RF alone and antifilaggrin antibody test was easy, convenient ELISA method in performance.

Diagnostic Performance of the Anti-Cyclic Citrullinated Peptide Antibodies in Rheumatoid Arthritis / 대한진단검사의학회지

BACKGROUND: The Rheumatoid Factor (RF) is the only serological marker in the diagnosis of rheumatoid arthritis (RA), but its sensitivity and specificity are not satisfactory for the diagnosis of RA. Therefore, we investigated the diagnostic performance of a new anti-cyclic citrullinated peptide antibodies test (anti-CCP) by the enzyme-linked immunosorbent assay (ELISA) in RA. METHODS: A cyclic peptide variant that contains citrulline was used as an antigenic substrate in ELISA. We performed the RF and anti-CCP in 324 RA patients, 251 non-RA patients (rheumatic diseases other than RA), and 286 normal individuals. Diagnostic performances such as sensitivity and specificity were evaluated by the receiver-operator characteristics (ROC) curve at optimal cut-off values. The optimal cut-off values were determined at the maximal point of the area under the curve. RESULTS: The sensitivity and specificity of anti-CCP were 72.8% and 92% at 3.8 U/mL. The sensitivity and specificity of RF were 80.6% and 78.5% at 9 U/mL. The sensitivity and specificity of anti-CCP and RF were 67%, 95.2% and 63.3%, 90% at 8.4 U/mL, 20 U/mL, respectively. A combination of anti-CCP with RF increased the sensitivity and specificity to 79.3%, 96.4%, respectively. Anti-CCP was positive in 23.8% among 63 sero-negative RA patients. CONCLUSIONS: We considered that the anti-CCP might be useful as another new serological marker for the diagnosis of a RA combination with RF, or not, because the anti-CCP has a higher diagnostic specificity than the RF and was an easy, convenient ELISA method in performance.

Analysis of Effectiveness of the Maximum Surgical Blood Order Schedule / 대한수혈학회지

BACKGROUND: The maximum surgical blood order schedule (MSBOS) is a viable option for reducing unnecessary crossmatching and achieving significant cost savings in the blood bank. In this study, we showed the process establishing MSBOS and through a prospective study, we evaluated the efficacy of MSBOS. METHODS: We organized task force team for transfusion management improvement composed of a director of the blood bank, surgeons and anesthesiologists in the Committee for Quality Improvement (CQI) of Asan Medical Center. In this team, we established MSBOS for most elective surgeries through the review of the previous transfusion and crossmatching data. We introduced MSBOS in April 1998 and prospectively analyzed surgeon's acceptance rate of MSBOS, crossmatch-to-transfusion ratio (C/T ratio), blood wastage rate, and cost savings. RESULTS: During the first 19 months after introducing MSBOS at our hospital, there was gradual increase in the surgeon's compliance rate of MSBOS from 30% to 94.0% through continuous education. The C/T ratio was changed from 3.5 to 1.6 and blood wastage rate was decreased from 4.0% to 1.9%. And also we could save more than 38,400,000 won through not performing the unnecessary crossmatches of 7,680 cases per year. CONCLUSION: Introduction of MSBOS can have a significant impact in reducing C/T ratio, blood wastage rate, and unnecessary crossmatches for the unused blood units. For successful application of MSBOS, cooperation with surgeons and anesthesiologists and continuous education is essential.

A Case of Identification of Marker Chromosome by comparative genomic hybridization and fluorescence in situ hybridization / 대한임상병리학회지

Comparative genomic hybridization (CGH) has been used to identify deletions and amplifications, particularly in neoplastic samples. CGH provides a new possibility searching genomes for imbalances of genetic material. We described the combined use of CGH and fluorescence in situ hybridization (FISH) to identify the origin of a marker chromosome in a child with mental retardation. Giemsa banding of metaphases from cultured lymphocytes showed a marker chromosome. The Karyotype was 47,XX,+mar. CGH revealed that the additional material originated from 15q. FISH confirmed this finding with whole chromosome paint for chromosome 15 and with a D15S10 (15q11-13) probe. This case demonstrates the efficient use of CGH and confirmatory FISH for the identification of chromosomal material of unknown origin.

Detailed Characterization of Chromosomal Breakpoints in an ALL, L3 using Cross Species Color Banding / 대한임상병리학회지

Karyotype analysis by G-anding is the standard method for identifying numerical and structural chromosomal aberrations in cytogenetic laboratories. However, the origins of marker chromosomes, subtle translocations, or complex chromosomal rearrangements are often difficult to identify with certainty. Cross-pecies color banding is a new FISH-ased screening technique that enables the generation of a specific color banding pattern for each human chromosome based on the genomic homologies between man and various species of apes. We report application of Cross-pecies color banding (RxFISH) to characterize the chromosomal rearrangements of one leukemia sample the G-and karyotype of which were incomplete. The combination of G-anding and RxFISH in this case study yielded additional information beyond that obtained by either technique used alone in determining the precise breakpoints in complex chromosomal rearrangements.

Detailed Characterization of Chromosomal Breakpoints in an ALL, L3 using Cross Species Color Banding / 대한임상병리학회지

Karyotype analysis by G-anding is the standard method for identifying numerical and structural chromosomal aberrations in cytogenetic laboratories. However, the origins of marker chromosomes, subtle translocations, or complex chromosomal rearrangements are often difficult to identify with certainty. Cross-pecies color banding is a new FISH-ased screening technique that enables the generation of a specific color banding pattern for each human chromosome based on the genomic homologies between man and various species of apes. We report application of Cross-pecies color banding (RxFISH) to characterize the chromosomal rearrangements of one leukemia sample the G-and karyotype of which were incomplete. The combination of G-anding and RxFISH in this case study yielded additional information beyond that obtained by either technique used alone in determining the precise breakpoints in complex chromosomal rearrangements.

Genetic Alterations in Bladder Cancer Detected by Comparative Genomic Hybridization / 대한암학회지

PURPOSE: Cytogenetic and genetic alterations of tumors are closely related with progressian and promotion of cancers. Comparative genomic hybridization (CGH) has known to be a novel tool for the detection of genetic alteration in solid cancers. We performed CGH for the detection of new genetic alterations of bladder tumors. MATERIALS AND METHODS: Biotin-labeled tumor DNA and digoxigenin-labeled normal DNA were hybridized to normal metaphase cells. The fluorescence signals were captured by fluorescence microscope after detection by avidin FITC and antidigoxigenin rhodamin. Then, the ratio of fluorescence was calculated by an image analyzer. RESULTS: CGH results showed amplifications on chromosomes 1q, 3q, 4q, 5p, 6pq, 7p, 8q, 11q, 12q, 13q, 17q, 18q and 20pq (more than 20% of cases). Deletions were on chromosome 2q21-qter, 4q13-q23, 5q, 8p12-p22, 9pq, 11p13-p15 (more than 20% of cases). High level amplifications were noted on chromosomes 1q31-qter, 3p21, 3q24, 4q26, Sq21-qter, llq14-qter, 12q15-q21, 12q21-q24, 13q21-q31, 17q22, 18q22. CONCLUSION: We considered that the amplification on chromosome 4q26, 11q14-qter, 12q21-q24, 18q12 and deletion on 4qll-4q13 as a novel genetic alterations of bladder cancer. Our results revealed different pattem of amplifications that affect other regions from previous study on chromosome 7, llq, 12q, 13q, and 18q. CGH was very useful for the screening of genetic alterations of solid tumors.

Detection of Helicobacter pylori using Polymerase Chain Reaction / 대한임상병리학회지

BACKGROUND: Helicobacter pylori(H. pylori) is an important etiologic agent for chronic active gastritis and plays a role in the pathogenesis of peptic ulcer and stomach cancer and recently lymphomas occurring in mucosa associated lymphatic tissue. At present, H. pylori infection associated gastritis was estimated about 80% among the cause of chronic gastritis. In this study, we tested Polymerase Chain Reaction (PCR) assay to detect H. pylori infection in gastric biopsy specimens. This results were compared with results obtained by other tests. METHODS: A total of 70 patients with dyspepsia were evaluated for H. pylori infection through the use of PCR, culture and serologic tests. The study population had an age of 12 to 80 years(median 46.4), there were 31 males and 39 females. We tested PCR using H. pylori detection kit(TM) (Bioneer, Korea) and anti-H. pylori anti-body EIA using GAP test IgG and IgM(TM)(BIO-RAD, USA). We used anaerobic jar without catalyst for the microaerophilic condition. RESULTS: The positive result by PCR assay for diagnosis of H. pylori infection in gastric specimens was 71.4% in total of 70 patients, which the gastritis, peptic ulcer and gastric cancer were 63.2%, 77.8% and 85.7%, respectively. Among 10 gastrectomy specimens of stomach cancers, the detection rate of H. pylori infection by culture was 50% and the PCR assay was 100%. The detection rate of If pylori IgG and IgM antibodies by commercially available GAP test IgG and IgM EIA were 64.3%, respectively, and IgG or IgM were 85.7%. CONCLUSIONS: The serologic study was sensitive but it was appeared that the high false positive (75%) and false negative (25%) rate and could not confirm current infection. The PCR assay was shown to be more sensitive, rapid and easy to treat specimen for the detection of H. pylori infection than conventional methods such as culture and serologic test in dyspeptic patients.

Detection of Helicobacter pylori using Polymerase Chain Reaction / 대한임상병리학회지

BACKGROUND: Helicobacter pylori(H. pylori) is an important etiologic agent for chronic active gastritis and plays a role in the pathogenesis of peptic ulcer and stomach cancer and recently lymphomas occurring in mucosa associated lymphatic tissue. At present, H. pylori infection associated gastritis was estimated about 80% among the cause of chronic gastritis. In this study, we tested Polymerase Chain Reaction (PCR) assay to detect H. pylori infection in gastric biopsy specimens. This results were compared with results obtained by other tests. METHODS: A total of 70 patients with dyspepsia were evaluated for H. pylori infection through the use of PCR, culture and serologic tests. The study population had an age of 12 to 80 years(median 46.4), there were 31 males and 39 females. We tested PCR using H. pylori detection kit(TM) (Bioneer, Korea) and anti-H. pylori anti-body EIA using GAP test IgG and IgM(TM)(BIO-RAD, USA). We used anaerobic jar without catalyst for the microaerophilic condition. RESULTS: The positive result by PCR assay for diagnosis of H. pylori infection in gastric specimens was 71.4% in total of 70 patients, which the gastritis, peptic ulcer and gastric cancer were 63.2%, 77.8% and 85.7%, respectively. Among 10 gastrectomy specimens of stomach cancers, the detection rate of H. pylori infection by culture was 50% and the PCR assay was 100%. The detection rate of If pylori IgG and IgM antibodies by commercially available GAP test IgG and IgM EIA were 64.3%, respectively, and IgG or IgM were 85.7%. CONCLUSIONS: The serologic study was sensitive but it was appeared that the high false positive (75%) and false negative (25%) rate and could not confirm current infection. The PCR assay was shown to be more sensitive, rapid and easy to treat specimen for the detection of H. pylori infection than conventional methods such as culture and serologic test in dyspeptic patients.