Current Status of Flow Cytometric Immunophenotyping of Hematolymphoid Neoplasms in Korea

Background@#Flow cytometric immunophenotyping of hematolymphoid neoplasms (FCIHLN) is essential for diagnosis, classification, and minimal residual disease (MRD) monitoring. FCI-HLN is typically performed using in-house protocols, raising the need for standardization. Therefore, we surveyed the current status of FCI-HLN in Korea to obtain fundamental data for quality improvement and standardization. @*Methods@#Eight university hospitals actively conducting FCI-HLN participated in our survey.We analyzed responses to a questionnaire that included inquiries regarding test items, reagent antibodies (RAs), fluorophores, sample amounts (SAs), reagent antibody amounts (RAAs), acquisition cell number (ACN), isotype control (IC) usage, positiveegative criteria, and reporting. @*Results@#Most hospitals used acute HLN, chronic HLN, plasma cell neoplasm (PCN), and MRD panels. The numbers of RAs were heterogeneous, with a maximum of 32, 26, 12, 14, and 10 antibodies used for acute HLN, chronic HLN, PCN, ALL-MRD, and multiple myeloma-MRD, respectively. The number of fluorophores ranged from 4 to 10. RAs, SAs, RAAs, and ACN were diverse. Most hospitals used a positive criterion of 20%, whereas one used 10% for acute and chronic HLN panels. Five hospitals used ICs for the negative criterion. Positiveegative assignments, percentages, and general opinions were commonly reported. In MRD reporting, the limit of detection and lower limit of quantification were included. @*Conclusions@#This is the first comprehensive study on the current status of FCI-HLN in Korea, confirming the high heterogeneity and complexity of FCI-HLN practices. Standardization of FCI-HLN is urgently needed. The findings provide a reference for establishing standard FCI-HLN guidelines.

Isolation of Carnobacterium divergens from Blood Culture in Korea : A Case Report and Literature Review / 대한임상미생물학회지

Carnobacterium is a genus of gram-positive bacilli belonging to the family Lactobacillaceae.Generally, Carnobacterium species are considered nonpathogenic to humans and are mostly found in the natural environment, food, and food packaging. Furthermore, some Carnobacterium species play a bioprotective role in the food industry. Isolation of Carnobacterium from human blood or other sites, such as skin or abscess, has rarely been reported—there are only four published case reports worldwide, and none of them is from Korea. In all the reported cases, the patients reported contact with an aqueous environment or were administered nutrition via a parenteral route. Herein, we report the detection of Carnobacterium divergens bacteremia in an immunocompromised patient by using mass spectrometry in Korea.

Evaluation of Automated Platelet Aggregation Test Using a Sysmex CS-5100 Analyzer

Background@#The platelet aggregation test is widely used to measure antiplatelet therapy response and to detect platelet function disorders. CS-5100 (Sysmex Co., Japan) is a recently introduced coagulation analyzer that can also measure platelet aggregation. We evaluated the performance of CS-5100 in the platelet aggregation test for use in clinical laboratories. @*Methods@#We investigated the precision, stability, dilution test, and correlation of CS-5100 with a traditional aggregometer. Precision was tested using normal and patient samples. Stability was assessed over 5 different time points for 8 hours. The dilution test was performed with normal samples using ADP agonists. We tested correlations between the results of Chrono-log aggregometer (Chrono-Log Co., USA) and CS-5100 using 10 samples with 5 agonists each. We also calculated the reference range of 5 agonists using 22-30 normal samples. @*Results@#The coefficients of variation using normal samples were 7.45% and 3.27% for ADP and arachidonic acid, respectively. Stability was maintained for up to 2 hours in most samples. Dilution tests showed similar results until reaching a dilution factor of 2. Strong correlations of the results between Chrono-log and CS-5100 were found, except for ristocetin. The reference ranges of 5 reagents in CS-5100 were similar to those obtained with the Chrono-log aggregometer. @*Conclusions@#The performance of CS-5100 in platelet aggregation tests showed reliable results compared to a traditional aggregometer. CS-5100 can perform coagulation test and platelet aggregation test, simultaneously. Thus, CS-5100 can enable cost saving and reduce turn-around-time by reducing the inspection time.

A Rare Case of Acute Myeloid Leukemia With SET-NUP214 Fusion and Massive Hyperdiploidy

No abstract available.

Recent progress in laboratory diagnosis of thalassemia and hemoglobinopathy: a study by the Korean Red Blood Cell Disorder Working Party of the Korean Society of Hematology

Genetic hemoglobin disorders are caused by mutations and/or deletions in the α-globin or β-globin genes. Thalassemia is caused by quantitative defects and hemoglobinopathies by structural defect of hemoglobin. The incidence of thalassemia and hemoglobinopathy is increased in Korea with rapid influx of people from endemic areas. Thus, the awareness of the disease is needed. α-thalassemias are caused by deletions in α-globin gene, while β-thalassemias are associated with decreased synthesis of β-globin due to β-globin gene mutations. Hemoglobinopathies involve structural defects in hemoglobin due to altered amino acid sequence in the α- or β-globin chains. When the patient is suspected with thalassemia/hemoglobinopathy from abnormal complete blood count findings and/or family history, the next step is detecting hemoglobin abnormality using electrophoresis methods including high performance liquid chromatography and mass spectrometry. The development of novel molecular genetic technologies, such as massively parallel sequencing, facilitates a more precise molecular diagnosis of thalassemia/hemoglobinopathy. Moreover, prenatal diagnosis using genetic testing enables the prevention of thalassemia birth and pregnancy complications. We aimed to review the spectrum and classification of thalassemia/hemoglobinopathy diseases and the diagnostic strategies including screening tests, molecular genetic tests, and prenatal diagnosis.

Recommendation for the Peripheral Blood Cell Morphology Report

There is considerable heterogeneity in the peripheral blood smear reports across different diagnostic laboratories, despite following the guidelines published by the International Council for Standardization in Haematology (ICSH). As standardization of reports can facilitate communication and consequently the diagnostic efficiency in both laboratories and clinics, the standardization committee of the Korean Society for Laboratory Hematology aimed to establish a detailed guideline for the standardization of peripheral blood smear reports. Based on the ICSH guidelines, additional issues on describing and grading the peripheral blood smear findings were discussed. In this report, the proposed guideline is briefly described.

Analytical Performance of New ARCHITECT AFP Assay: Comparison with the Current Assay

Alpha-fetoprotein (AFP) is frequently used for hepatocellular carcinoma (HCC) diagnosis and surveillance. Although the current ARCHITECT AFP (List number 7K67) assay range is 0–350 ng/mL, all samples with test results between 200 and 350 ng/mL must be diluted and retested until their levels are <200 ng/mL. A new ARCHITECT AFP (8100/3P36) assay with a dynamic range of up to 2000 ng/mL has been introduced. The aim of this study was to perform a method comparison between the current ARCHITECT AFP assay and the new assay. The precision study showed excellent results for both high and low controls. There was a positive correlation between the two assay systems and clinical samples. The new ARCHITECT AFP assay with a wide assay range demonstrated good analytical performance. Therefore, the current ARCHITECT AFP assay could be replaced by the new assay, which is more convenient and minimizes manual labor.

Platelet Function Analyzer-200 P2Y Results Are Predictive of the Risk of Major Adverse Cardiac Events in Korean Patients Receiving Clopidogrel Therapy Following Acute Coronary Syndrome

BACKGROUND: Clopidogrel is one of the most commonly used anti-platelet agents in cardiovascular diseases. We analyzed the relationship between the platelet function analyzer (PFA)-200 P2Y (INNOVANCE PFA-200 System, Siemens Healthcare, Germany) results and occurrence of major adverse cardiac events (MACEs) in Korean patients with recent-onset acute coronary syndrome (ACS) taking clopidogrel. METHODS: Between August 2013 and June 2016, we prospectively enrolled 106 patients with recent-onset ACS who had been treated with clopidogrel. We obtained blood samples and measured closure time (CT) using the PFA-200 P2Y test. Patients were divided into two groups on the basis of a CT cut-off value of 106 seconds. We compared patient characteristics and various MACEs that occurred during the follow-up period. RESULTS: The CTs for 78 patients exceeded the cut-off value. At the time of these analyses, 11 patients had been diagnosed with MACEs. In the time-to-event analysis, there was a difference between the two groups (P<0.001). After adjusting other variables associated with MACE occurrence, CT value was the strongest predictor of MACEs, with a 7.30-fold occurrence risk (P=0.002). CONCLUSIONS: We found a strong relationship between CT and MACE risk in Korean patients with recent-onset ACS taking clopidogrel. Accordingly, PFA-200 P2Y results could be used as a predictive marker for MACE risk in such patients.

Philadelphia-positive mixed phenotype acute leukemia presenting with PML-RARα fusion transcript without t(15;17) on cytogenetic studies

No abstract available.

A Case of Bone Marrow Necrosis in B-Acute Lymphoblastic Leukemia with Philadelphia Chromosome at Presentation

Bone marrow necrosis (BMN) is a pathologic state which is derived from various disease entities. Most commonly, it is accompanied by hematologic malignancies such as acute leukemia. The patients with marrow necrosis are generally known to have dismal prognoses but variations exist according to early diagnosis. Here we report a case of BMN in an acute lymphoblastic leukemia patient with Philadelphia chromosome at presentation.

A Case with Changes in Autoantibodies with Mimicking Specificity in a Patient with Hepatocellular Carcinoma / 대한수혈학회지

Changes in autoantibodies with mimicking specificity for Rh antigens are quite rare. Here, we report a 73-year-old male patient with hepatocellular carcinoma that showed changes in the autoantibodies with mimicking specificity. He was admitted to our hospital with mental alterations and hypoglycemia. The antibody identification test for RBC transfusion showed a pattern of agglutination, suggesting anti-E alloantibody and anti-c autoantibody. The patient was then diagnosed with hepatocellular carcinoma, and after 18 months, the antibody identification test showed a different pattern of agglutination, suggesting anti-E alloantibody and anti-C autoantibody. We recommend a careful evaluation to make sure an appropriate transfusion is performed when encountering fluctuating autoantibodies with mimicking specificity.

NUP98 Rearrangement in Acute Myelomonocytic Leukemia With t(11;19)(p15;p12): The First Case Report Worldwide

No abstract available.

A Case of Acute Promyelocytic Leukemia with Co-existence of BCR-ABL1 and PML-RARA Rearrangements Detected by PCR

Acute promyelocytic leukemia (APL) is a distinct subtype of acute myeloid leukemia (AML) characterized by PML-RARA fusion and specific morphology. The BCR-ABL1 rearrangement is mainly observed in patients with chronic myeloid leukiemia (CML). However, it is also found in patients with acute lymphoblastic leukemia (ALL) and in a few patients with AML. However, it is very rarely observed in patients with APL. Here, we report a case of APL with t(15;17) and co-existence of PML-RARA and BCR-ABL1. Further study with more cases is warranted to find the right treatment and prognostic significance.

Revisiting the Pre-transfusion Test: A Case of Acute Hemolytic Transfusion Reaction due to Multiple Alloantibodies of Anti-E, Anti-c, Anti-Jk(b) / 대한수혈학회지

We report a case of acute hemolytic transfusion reaction due to multiple alloantibodies. A 41-year-old male with multiple histories of transfusion was admitted for jaundice and oliguria after receiving two units of red blood cells in a local clinic. He showed acute renal failure and disseminated intravascular coagulation. Direct Coombs test was negative and antibody screening test showed strong positive results. Anti-E, anti-c, and anti-Jk(b) antibodies were identified in two panels of unexpected antibody assays. Acute hemolytic transfusion was diagnosed, and he was discharged after 1 month of supportive treatment. Unexpected antibody detection tests, including the antiglobulin phase, should be performed to prevent adverse transfusion reactions by unexpected antibodies. Better precision and quality control are necessary when performing pre-transfusion tests.

Relationship Between Cytogenetic Complexity and Peritumoral Edema in High-Grade Astrocytoma

BACKGROUND: The purpose of the study is to reveal the association of cytogenetic compltyexi and peritumoral edema volume (PTEV) and its prognostic significance in high-grade astrocytoma patients by culturing patient tumor cells. METHODS: Twenty-seven high-grade astrocytoma patients were divided into three groups according to karyotype complexity: normal, non-complex karyotype (NCK), and complex karyotype (CK). Endothelial growth factor receptor (EGFR) amplification was detected by FISH, and its association with chromosome 7 abnormalities was analyzed. Mean PTEV of each group was compared by ANOVA to evaluate the relationship between PTEV and cytogenetic complexity. RESULTS: The PTEV of patients in normal (n=6), NCK (n=8), and CK (n=13) groups were 24.52±17.73, 34.26±35.04, and 86.31±48.7 cm3, respectively (P=0.005). Ten out of 11 patients with EGFR amplification showed abnormalities in chromosome 7. The mean PTEV of EGFR-amplified and non-amplified groups were 80.4±53.7 and 41.3±37.9 cm3, respectively (P=0.035). The average survival of patients with PTEV less than 90 cm3 was 30.52±26.11 months, while in patients with PTEVs over or equal to 90 cm3, it was 10.83±5.53 months (P=0.007). CONCLUSIONS: The results show an association of complex karyotype with the PTEV of high-grade astrocytoma. EGFR amplification plays a significant role in the formation of peritumoral edema, causing PTEV to increase, which is related with survival. This implies that cytogenetic karyotype can be applied as a prognostic factor.

A retrospective analysis of cytogenetic alterations in patients with newly diagnosed multiple myeloma: a single center study in Korea

BACKGROUND: The accurate identification of cytogenetic abnormalities in multiple myeloma (MM) has become more important over recent years for the development of new diagnostic and prognostic markers. In this study, we retrospectively analyzed the cytogenetic aberrations in MM cases as an initial assessment in a single institute. METHODS: We reviewed the cytogenetic results from 222 patients who were newly diagnosed with MM between January 2000 and December 2015. Chromosomal analysis was performed on cultured bone marrow samples by standard G-banding technique. At least 20 metaphase cells were analyzed for karyotyping. RESULTS: Clonal chromosome abnormalities were detected in 45.0% (100/222) of the patients. Among these results, 80 cases (80.0%) had both numerical and structural chromosome abnormalities. Overall hyperdiploidy with structural cytogenetic aberrations was the most common finding (44.0%), followed by hypodiploidy with structural aberrations (28.0%). Amplification of the long arm of chromosome 1 and -13/del(13q) were the most frequent recurrent abnormalities, and were detected in 50 patients (50.0%) and 40 patients (40.0%) with clonal abnormalities, respectively. The most common abnormality involving 14q32 was t(11;14)(q13;q32), which was observed in 19 cases. CONCLUSION: These findings demonstrate that myeloma cells exhibit complex aberrations regardless of ploidy, even from a single center in Korea. Conventional cytogenetic analysis should be included in the initial diagnostic work-up for patients suspected of having MM.

Repeatability of QuantiFERON-TB Gold In-Tube Assay Results Near Cut-Off Points

No abstract available.

Development and Characterization of Reference Materials for Genetic Testing: Focus on Public Partnerships

Characterized reference materials (RMs) are needed for clinical laboratory test development and validation, quality control procedures, and proficiency testing to assure their quality. In this article, we review the development and characterization of RMs for clinical molecular genetic tests. We describe various types of RMs and how to access and utilize them, especially focusing on the Genetic Testing Reference Materials Coordination Program (Get-RM) and the Genome in a Bottle (GIAB) Consortium. This review also reinforces the need for collaborative efforts in the clinical genetic testing community to develop additional RMs.

A Rare der(Y)t(Y;1)(q12;q12) in a Patient with Essential Thrombocythemia

Among hematologic diseases, structural abnormalities of autosomal chromosomes are well-known, but cases involving the sex chromosomes are uncommon. Duplications of the long arm of chromosome 1 have been reported in several hematologic diseases including myelodysplastic syndrome, myeloproliferative neoplasms, acute myeloid leukemia, acute lymphoblastic leukemia, and Burkitt lymphoma. However, dup(1q) as a der(Y)t(Y;1)(q12;q12) is very rare. Here, we report a case of essential thrombocythemia harboring der(Y)t(Y;1)(q12;q12) with literature review.

Correlation Between the CYP2C19 Phenotype Status and the Results of Three Different Platelet Function Tests in Cardiovascular Disease Patients Receiving Antiplatelet Therapy: An Emphasis on Newly Introduced Platelet Function Analyzer-200 P2Y Test

BACKGROUND: An association has been reported between CYP2C19 polymorphism and the altered antiplatelet activity of clopidogrel. We investigated this association using the newly introduced platelet function analyzer (PFA)-200 (INNOVANCE PFA-200 System; Siemens Healthcare, Germany) P2Y test. METHODS: Polymorphisms of CYP2C19*2, *3, *17 and the degree of inhibition of platelet function were determined in 83 patients. Three different platelet function tests were used to evaluate the degree of platelet inhibition and to check the association with genotype. RESULTS: The post-procedure PFA-200 values of extensive metabolizers (EM) patients (285.3+/-38.8) were higher than those of intermediate metabolizers (IM) and poor metabolizers (PM) patients (227.7+/-98.3 and 133.7+/-99.2, respectively; P=0.024). Light transmittance aggregometry (LTA) and the VerifyNow system showed that the post-procedure values for EM patients were lower than those of IM and PM patients (LTA: 24.4+/-15.7, 34.1+/-17.6, and 42.2+/-16.9, respectively, P<0.001; VerifyNow: 133.2+/-60.5, 171.5+/-42.6, and 218.7+/-59.3, respectively, P<0.001). The high residual platelet reactivity (HPR) rates were significantly different among the EM, IM, and PM groups using PFA-200 (PM:IM:EM=82.4:40.6:11.8, P<0.001). CONCLUSIONS: Approximately, 59.0% of Korean patients with cardiovascular disease receiving clopidogrel had CYP2C19 loss-of-function genotypes classified as IM or PM, and the frequency was similar to the data from Asian people. The PFA-200, LTA, and VerifyNow platelet function tests revealed evidence of a significant association between the efficacy of clopidogrel and CYP2C19 genotypes.