The Frequency and Distribution of Unexpected Antibodies in Transfusion Candidates with the Use of the Ortho BioVue System: Recent Four Year Experience / 대한수혈학회지

BACKGROUND: Unexpected antibodies are important factors for hemolytic transfusion reactions. In the past, the tube method was used for detecting unexpected antibodies. The column agglutination method has recently been widely used because of its simplicity and it has a higher rate of detecting warm antibodies. In this study, we describe the frequency and distribution of unexpected antibodies in transfusion candidates during the recent 4 years and the transfusion characteristics in the identified cases. METHODS: Antibody screening tests were carried out on 44,008 sera using the column agglutination method from January, 2005 to December, 2008. The antibodies were screened and identified by the Ortho BioVue System (Ortho-Clinical Diagnostics, Raritan, NJ, USA). RESULTS: Of the 44,008 cases that underwent unexpected antibodies screening, 589 cases (1.3%) showed positive results. Unexpected antibodies were identified in 383 cases. The antibodies that were most frequently detected were anti-Lewis antibodies in 130 cases (34.0%). Among the warm antibodies, anti-Rh and anti-Kidd antibodies were detected in 67 cases (17.5%) and 2 cases (0.5%), respectively. Unidentified antibodies were detected in 133 cases (38.9%). Among the patients with unexpected antibodies, 137 cases (35.8%) had a history of previous transfusion and 244 cases (63.7%) had a history of previous transfusion or gestation. CONCLUSION: Anti-Lewis cold antibodies were the most frequently detected antibodies. Warm antibodies were also frequently detected, and these are clinically significant.

Utility of In-House PCR for HLA-B27 Typing: Comparison of Concordance Rate between PCR Kit and In-House PCR / 대한진단검사의학회지

BACKGROUND: Commercial kits of PCR method are widely used in HLA-B27 typing; however, their cost is relatively high. In this study, we evaluated the utility of an in-house PCR method by comparing it with that of a commercial kit. METHODS: HLA-B27 typing was done in 188 patients by using two PCR methods, Absolute(TM) HLAB27 PCR kit (Biosewoom, Korea) and an in-house PCR method. The primers used in the in-house method were prepared by Bioneer (Korea). Both PCR tests were done by Gene Amp PCR System 9600 (Perkin-Elmer Centus Corp., USA). RESULTS: The commercial kit and in-house PCR showed 100% concordance rate with each other in HLA-B27 typing. Of 188 patients tested 72 (38.3%) were positive and 116 (61.7%) were negative by the both tests. Of 62 patients with ankylosing spondylitis, 50 were positive (80.7%). CONCLUSIONS: The in-house PCR is a reliable and cost-effective method and can replace or supplement commercial kits for HLA-B27 typing.

Evaluation of CELL-DYN Sapphire Hematology Analyzer / 임상검사와정도관리

BACKGROUND: CELL-DYN Sapphire (Abbott Diagnostics, Santa Clara, CA, USA) hematology analyzer is based on laser multi angle polarized scatter separation, focused flow impedance and fluorescent flow cytometry. In order to assess the usefulness of this newly introduced analyzer, we evaluated the precision and correlation of complete blood count including white blood cell differential count and reticulocyte count of CELL-DYN Sapphire instrument. METHODS: Patient samples ordered for complete blood count and white blood cell differential count and CELL-DYN 29 Plus control samples were used. We evaluated the precision of complete blood count and WBC differential count and analyzed correlation of these parameters for comparison with ADVIA 120 (Bayer corporation, Tarrytown, NY, USA) and manual count. We additionally evaluated reticulocyte parameters for precision and comparison with ADVIA 120. RESULTS: Short term precisions showed low coefficient of variation (CV) values for all CBC parameters including lower than 3% CV values for neutrophil and lymphocyte count. Long term precision showed lower than 4% CV values for all CBC and WBC differential count except for basophils. Comparison with ADVIA 120 showed good correlation for CBC except for MCHC. Good correlation was confirmed in all differential counts except for basophils. Comparison with manual WBC differential count showed good correlation except for monocyte and eosinophil. CONCLUSIONS: CELL-DYN Sapphire has high precision and good correlation with ADVIA 120 and manual count. CELL-DYN Sapphire is an exceptional instrument for CBC with WBC differentials and reticulocyte counting.

Evaluation of CELL-DYN Sapphire Hematology Analyzer / 임상검사와정도관리

BACKGROUND: CELL-DYN Sapphire (Abbott Diagnostics, Santa Clara, CA, USA) hematology analyzer is based on laser multi angle polarized scatter separation, focused flow impedance and fluorescent flow cytometry. In order to assess the usefulness of this newly introduced analyzer, we evaluated the precision and correlation of complete blood count including white blood cell differential count and reticulocyte count of CELL-DYN Sapphire instrument. METHODS: Patient samples ordered for complete blood count and white blood cell differential count and CELL-DYN 29 Plus control samples were used. We evaluated the precision of complete blood count and WBC differential count and analyzed correlation of these parameters for comparison with ADVIA 120 (Bayer corporation, Tarrytown, NY, USA) and manual count. We additionally evaluated reticulocyte parameters for precision and comparison with ADVIA 120. RESULTS: Short term precisions showed low coefficient of variation (CV) values for all CBC parameters including lower than 3% CV values for neutrophil and lymphocyte count. Long term precision showed lower than 4% CV values for all CBC and WBC differential count except for basophils. Comparison with ADVIA 120 showed good correlation for CBC except for MCHC. Good correlation was confirmed in all differential counts except for basophils. Comparison with manual WBC differential count showed good correlation except for monocyte and eosinophil. CONCLUSIONS: CELL-DYN Sapphire has high precision and good correlation with ADVIA 120 and manual count. CELL-DYN Sapphire is an exceptional instrument for CBC with WBC differentials and reticulocyte counting.

Inflammatory Marker Expression and Its Implication in Korean Ischemic Stroke Patients / 대한진단검사의학회지

BACKGROUND: Ischemic stroke is a complex condition influenced by many factors. Previous studies have demonstrated that inflammatory markers might play a role in such vascular diseases. Therefore the purpose of this study was to compare the expression of inflammatory markers in Korean ischemic stroke patients and to investigate their relationship to APOE polymorphism. METHODS: The patient group consisted of 275 patients with large artery atherosclerosis (LAA, n=169) and small artery occlusion (SAO, n=106). One hundred and nineteen age matched healthy subjects were recruited as the control group. Serum levels of three inflammatory markers (matrix metalloproteinase, MMP-9; tissue inhibitor of metalloproteinase-1, TIMP-1; and high-sensitivity C-reactive protein, hs-CRP) were measured in each patient by using commercially available kits. Comparison of clinical risk factors, inflammatory marker levels, and APOE genotypes between the stroke patient group and control group and between the two patient subgroups was assessed. RESULTS: Comparison of the stroke group to control group showed significantly elevated levels of circulating MMP-9 (P<0.01) and hs-CRP (P=0.01). Comparison between the individual subgroups revealed a significantly higher level of only TIMP-1 in the LAA subgroup compared to the SAO subgroup (P<0.01). There was no significant difference in inflammatory marker levels among each allele carrier. CONCLUSIONS: The present study revealed the obvious tendency of increased circulating inflammatory markers in the patients with acute ischemic attack, especially MMP-9 and hs-CRP. Our observations suggest that measurement of serum MMP-9, TIMP-1, and hs-CRP levels may be useful in the diagnosis of ischemic stroke patients.

A Case of Unspecified Mature T-cell Leukemia with Clover-shaped, Multi-lobated Nuclei

Mature T-cell leukemias are a group of neoplasms derived from mature or post-thymic T-cells, and a number of distinctive disease entities have been defined in the World Health Organization (WHO) classification. Here we report a 54-year-old female patient with multi-lobated atypical cells expressing the classic T-cell antigens involving multiple lymph nodes, peripheral blood, and bone marrow. The clinical, laboratory, and pathologic features of her disease did not fit into any of the entities in the WHO Classification. There was no evidence of rapidly rising lymphocyte counts, TCL1 expression, eosinophilia, erythroderma, Sezary cells, autoimmune phenomena, cytotoxic granules, nor evidence of HTLV-1 infection, and thus, T-cell prolymphocytic leukemia, Sezary syndrome, T-cell granular lymphocytic leukemia, and adult T-cell leukemia/lymphoma were all ruled out. This case suggests that further characterization and definition of the "unclassifiable" cases of mature T-cell neoplasm is needed to better understand the group of disorders.

Investigation of Hemostatic Changes in Patients with Sepsis / 대한진단검사의학회지

BACKGROUND: It is known that severe infection and inflammation lead to hemostatic abnormalities. Recently, much attention is focused on the mechanisms of infection or inflammation and on how it plays a central role in effecting the coagulation system. Disseminated intravascular coagulation in particular, is a common phenomenon in patients with sepsis, but the clinical implications of this condition are not clear. Therefore we attempted to evaluate the changes of the coagulation system in patients with sepsis and studied the factors that lead to such changes. METHODS: One hundred one patients diagnosed with sepsis were enrolled in this study. The patients were clinically evaluated for underlying disease and data for inflammatory status and coagulative changes were evaluated retrospectively. RESULTS: The WBC count increased in 76% and decreased in 6% of sepsis patients in comparison to the reference interval. The platelet count decreased in 65.3%. Changes in coagulative tests such as prothrombin time, activated partial thromboplastin time, antithrombin III, and D-dimer were observed in 70.4%, 52.7%, 87.2% and 100% of the patients, respectively. Correlation between ESR and fibrinogen was the highest in relation to the other coagulation factors. CRP also showed the highest correlation with fibrinogen in contrast to the other coagulation factors. CONCLUSIONS: This study confirmed the clear activation of coagulation in patients with sepsis. Of the evaluated factors involved in coagulation and fibrinolysis, fibrinogen showed the highest correlation to indices representing the inflammatory state. However further studies on the anticoagulant pathway are necessary in elucidating this matter.

A Case of Bacteremia Caused by Leuconostoc lactis Identified by 16S rRNA Sequencing / 대한임상미생물학회지

Leuconostoc spp. used to be considered nonpathogens to human, but human infections have been reported including bacteremia in compromized patients. Leuconostoc spp. may be misidentified as lactobacilli, streptococci, pediococci and enterococci due to atypical biochemical tests. Leuconostoc spp. are intrinsically resistant to vancomycin. We report a case of bacteremia caused by Leuconostoc lactis identified by 16S rRNA sequencing; the isolate was not identified by commercial kits.

Comparison of In-house Polymerase Chain Reaction Assay with Conventional Techniques for the Detection of Mycobacterium tuberculosis / 대한진단검사의학회지

BACKGROUND: The polymerase chain reaction (PCR) assay, introduced as a fast and sensitive diagnostic method, has been known to be useful in detecting Mycobacterium tuberculosis. Therefore the purpose of this study was to evaluate the usefulness of an in-house PCR assay in the detection of Mycobacterium tuberculosis by comparing PCR results with those of conventional diagnostic techniques. METHODS: We assessed the diagnostic yield of the in-house PCR assay retrospectively based on the patient's medical records using data from previously evaluated specimens submitted for PCR amplification IS6110 sequences by GeneAmp PCR system 9600 (Perkin Elmer, CT, USA). All samples had been examined for detection of M. tuberculosis by acid-fast stain and culture assay and the results from the 3 methods were analyzed. RESULTS: The majority of cases (1,727 cases, 96.6%) showed concordant results between in-house PCR, AFB stain, and culture methods; only 60 cases (3.4%) displayed discordant results. The sensitivities, specificities and positive and negative predictive values of each method were as follows: 81.0%, 99.6%, 95.0% and 98.4%, respectively for the in-house PCR; 63.4%, 100%, 100% and 96.9%, respectively for AFB staining method; and 83.8%, 100%, 100% and 98.6%, respectively for culture assays. CONCLUSIONS: The PCR assay shows a high sensitivity and specificity and is a reliable test for an early diagnosis of tuberculosis.

Usefulness of Anti-HCV ELISA Test and HCV Reverse Transcriptase-PCR for the Diagnosis of Hepatits C Viral Infection / 대한진단검사의학회지

BACKGROUND: The diagnosis of hepatitis C virus (HCV) infection is screened by anti-HCV enzymelinked immunosorbant assay (ELISA) and confirmed by recombinant immunoblotting assay (RIBA) or HCV RT-PCR. We attempted to evaluate the results between anti-HCV ELISA and a qualitative HCV RT-PCR. METHODS: Four hundred and twenty patients who were tested with anti-HCV ELISA and HCV RTPCR, simultaneously, from January 2002 to June 2005 were enrolled in this study. Anti-HCV ELISA was performed by AxSYM HCV version 3.0 (Abbott Laboratories, USA). HCV RT-PCR was performed using in-house RT-nested PCR methods from January 2002 to October 2004 and HCV Genotype Amplification Kit (LiPA) (Bayer Healthcare, USA) from November 2004 to June 2005. RESULTS: Of the 420 patients tested, 321 were positive for anti-HCV ELISA, and 204 were positive for RT-PCR. The positive predictability of anti-HCV ELISA was 63.6%. Among anti-HCV positive patients, RT-PCR was positive in 7.3% of the patients with sample/cut-off (S/CO) or =6. Among the 117 patients with positive anti-HCV, but with negative HCV RT-PCR, 64 had liver diseases such as chronic hepatitis C, chronic hepatitis B, or hepatocellular carcinoma. Twelve patients showed positive HCV RT-PCR, but negative anti-HCV results; of these 9 had hepatic dysfunction. CONCLUSIONS: In the patients who were positive for anti-HCV ELISA with a low S/CO, HCV RT-PCR positivity was shown in a low proportion. Therefore, in such cases, the results should be confirmed by RIBA or HCV RT-PCR. The liver function test showed increased levels of hepatic enzymes in patients with positive HCV RT-PCR, but negative anti-HCV. Such findings correlate to an early phase of chronic hepatitis C, suggesting the necessity of continuous follow up.

A Case of Acute Lymphoblastic Leukemia with Maturation

Acute lymphoblastic leukemia with maturation (ALLm) has different disease characteristics that does typical ALL. ALLm is characterized by an increased number of mature appearing leukemic cells (>20% of ANCs in the BM) having differentiation beyond the prolymphocyte stage according to light microscopic (LM) examination. It also has a worse prognosis than typical ALL. We have recently experienced a case of ALLm and we report on this case along with a literature review. A 36 year old patient showed lymphoblasts and mature appearing leukemic cells that were counted up to 15.8% and 23.0%, respectively, of the WBCs on bone marrow examination. Despite their mature appearance, these cells showed positivity for Tdt, CD10, CD19 and HLA-DR on the immunophenotypic study. Differentiating the mature-appearing leukemic cells from the hematogones or mature lymphocytes is difficult, and only through immunophenotypic examination is it possible to discriminate ALLm from typical ALL. We suggest performing a leukemic marker study that includes CD38 to effectively differentiate mature appearing leukemic cells from hematogones, especially for the follow up of leukemia with mature appearing cells.