Molecular Description of Macroorchis spinulosus (Digenea: Nanophyetidae) Based on ITS1 Sequences

We performed a molecular genetic study on the sequences of 18S ribosomal RNA (ITS1 region) gene in 4-day-old adult worms of Macroorchis spinulosus recovered in mice experimentally infected with metacercariae from crayfish in Jeollanam-do Province, Korea. The metacercariae were round, 180 µm in average diameter, encysted with 2 layers of thick walls, but the stylet on the oral sucker was not clearly seen. The adult flukes were oval shape, and 760-820 µm long and 320-450 µm wide, with anterolateral location of 2 large testes. The phylogenetic tree based on ITS1 sequences of 6 M. spinulosus samples showed their distinguished position from other trematode species in GenBank. The most closely resembled group was Paragonimus spp. which also take crayfish or crabs as the second intermediate host. The present study is the first molecular characterization of M. spinulosus and provided a basis for further phylogenetic studies to compare with other trematode fauna in Korea.

Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based VITEK MS System for the Identification of Acinetobacter Species from Blood Cultures: Comparison with VITEK 2 and MicroScan Systems

BACKGROUND: Acinetobacter species are the leading cause of bloodstream infection (BSI), but their correct identification is challenging. We evaluated the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based VITEK MS (bioMerieux, France), and two automated systems, VITEK 2 (bioMerieux) and MicroScan (Siemens, USA) for identification of Acinetobacter BSI isolates. METHODS: A total of 187 BSI isolates recovered at a university hospital in Korea between 2010 and 2012 were analyzed. The identification results obtained using VITEK MS and two automated systems were compared with those of rpoB sequencing. RESULTS: Of 187 isolates analyzed, 176 were identified to the species level by rpoB sequencing: the Acinetobacter baumannii group (ABG; 101 A. baumannii, 43 A. nosocomialis, 10 A. pittii isolates) was most commonly identified (82.4%), followed by Acinetobacter genomic species 13BJ/14TU (5.3%), A. ursingii (2.1%), A. soli (2.1%), A. bereziniae (1.1%), and A. junii (1.1%). Correct identification rates to the species group (ABG) level or the species level was comparable among the three systems (VITEK MS, 90.3%; VITEK 2, 89.2%; MicroScan, 86.9%). However, VITEK MS generated fewer misidentifications (0.6%) than VITEK 2 (10.8%) and MicroScan (13.1%) (P<0.001). In addition, VITEK MS demonstrated higher specificity (100%) for discrimination between ABG and non-ABG isolates than the other systems (both, 31.8%) (P<0.001). CONCLUSIONS: The VITEK MS system is superior to the VITEK 2 and MicroScan systems for identification of Acinetobacter BSI isolates, with fewer misidentifications and better discrimination between the ABG and non-ABG isolates.

Comparison of FcRgamma-Deficient and CD57+ Natural Killer Cells Between Cord Blood and Adult Blood in the Cytomegalovirus-Endemic Korean Population

BACKGROUND: FcRgamma-deficient natural killer (NK) cells (g-NK cells) have been associated with cytomegalovirus (CMV) infection. However, the frequency of g-NK cells in a CMV-endemic area (i.e., Korea) has not yet been studied. We examined the frequency of g-NK cells and expression of CD57 on NK cells in cord blood (CB) and adult blood (AB). METHODS: Of the 24 AB samples collected, 95.8% (23/24) were CMV IgG+/IgM-, while 100% of the 13 healthy CB samples were CMV IgG+/IgM-. We performed whole-blood flow cytometry assays to analyze intracellular FcRgamma and CD3zeta expression of CD3-/CD56dim NK cells from 13 CB and 24 AB samples, and surface CD57 expression on CD3-/CD56dim/CD16+ NK cells from 13 CB and 19 AB samples. RESULTS: All CMV seropositive AB samples contained g-NK cells (23/23), and the median proportion of g-NK cells in the CD3-/CD56dim NK cell pool was 35.0% (range: 11-77%). CD57+ NK cells in the CD3-/CD56dim/CD16+ NK cell population were detected in all 19 AB samples tested, but not in any CB samples. CONCLUSIONS: Our data suggest that g-NK cells and CD57+ NK cells are present at a very high frequency in CMV-seropositive AB, but rare in CMV-naive CB.

Evaluation of Modified Formalin-Ether Concentration Method Using Para Tube in Clinical Settings

Conventional formalin-ether concentration method is a gold standard for the diagnosis of parasite infection. However, it may be time-consuming and laborious. We aimed to reveal the clinical usefulness of a modified formalin-ether concentration method using the Para Tube (KS Corporation, Korea) compared with the conventional method. A total of 117 fresh, unpreserved fecal samples composed to 90 negative controls and 27 positive controls with ova of Diphyllobothrium latum/D. nihonkaiense, ova of Clonorchis sinensis and cysts of Giardia lamblia were used in this study. Both methods showed comparable correct identification rate (87.2% for the Para Tube vs. 86.3% for the conventional method).When five samples were examined at once, the Para Tube method reduced the procedure time compared with the conventional method (19 min 58 sec vs. 23 min 18 sec, P=0.0286). We concluded that the modified formalin-ether concentration method using the Para Tube is a rapid, simple, and reliable fecal concentration method for clinical use.

Seroepidemiology of Toxocariasis and Its Clinical Implications in Gwangju and Jeonnam-province, Korea

We investigated the seroepidemiological, clinical, and laboratory characteristics of patients suspected to have toxocariasis in Gwangju and Jeonnam-province, Korea. In total, 228 specimens were analyzed for anti-Toxocara canis IgG at two university hospitals from 2010 to 2012. The overall seropositive rate was 67.1%, and the seropositive rates among the eosinophilic and non-eosinophilic groups were 76.1% (105/138) and 53.3% (48/90), respectively. Risk factors for eosinophilia and toxocariasis were male sex (odds ratios [OR]=2.632 and 3.477, respectively) and a history of ingesting raw meat (OR=2.884 and 3.274, respectively), especially raw cow liver (OR=2.089 and 10.038, respectively). T. canis seropositivity (OR=5.807, P=0.004) and a history of consuming raw cow liver (OR=2.766, P=0.052) were risk factors for organ involvement. The anti-T. canis IgG level showed weakly positive correlations with eosinophil counts (r=0.234, P<0.001) and the duration of eosinophilia (r=0.155, P=0.019). Although limited to the regions of Gwangju and Jeonnam-province, this study supports the opinion that toxocariasis is a reasonable focus as a cause of eosinophilia and that it is also associated with organ involvement.

The cis-AB01 Allele Originated From the A105 Allele, and not From the A102 Allele

No abstract available.

Comparison of AdvanSure TB/NTM PCR and COBAS TaqMan MTB PCR for Detection of Mycobacterium tuberculosis Complex in Routine Clinical Practice

The AdvanSure tuberculosis/non-tuberculous mycobacterium (TB/NTM) PCR (LG Life Science, Korea) and COBAS TaqMan Mycobacterium tuberculosis (MTB) PCR (Roche Diagnostics, USA) are commonly used in clinical microbiology laboratories. We aimed to evaluate these two commercial real-time PCR assays for detection of MTB in a large set of clinical samples over a two-year period. AdvanSure TB/NTM PCR and COBAS TaqMan MTB PCR were performed on 9,119 (75.2%) and 3,010 (24.8%) of 12,129 (9,728 respiratory and 2,401 non-respiratory) MTB specimens, with 361 (4.0%) and 102 (3.4%) acid-fast bacilli (AFB)-positive results, respectively. In MTB culture, 788 (6.5%) MTB and 514 (4.2%) NTM were identified. The total sensitivity and specificity of the AdvanSure assay were 67.8% (95% confidence interval [CI], 63.9-71.6) and 98.3% (95% CI, 98.0-98.6), while those of the COBAS TaqMan assay were 67.2% (95% CI, 60.0-73.8) and 98.4% (95% CI, 97.9-98.9), respectively. The sensitivities and specificities of the AdvanSure and COBAS TaqMan assays for AFB-positive and AFB-negative samples were comparable. Furthermore, the AdvanSure assay showed fewer invalid results compared with the COBAS TaqMan assay (5.0 vs. 20.4 invalid results/1,000 tests, P<0.001). AdvanSure assay represents a comparable yet more reliable method than COBAS TaqMan for the identification of mycobacteria in routine clinical microbiology.

Cryptic e1a2 BCR-ABL1 Fusion With Complex Chromosomal Abnormality in de novo Myelodysplastic Syndrome

No abstract available.

Strongyloidiasis in a Diabetic Patient Accompanied by Gastrointestinal Stromal Tumor: Cause of Eosinophilia Unresponsive to Steroid Therapy

We report here a case of strongyloidiasis in a 72-year-old diabetic patient (woman) accompanied by gastrointestinal stromal tumor receiving imatinib therapy, first diagnosed as hypereosinophilic syndrome and treated with steroids for uncontrolled eosinophilia. She suffered from lower back pain and intermittent abdominal discomfort with nausea and diagnosed with gastrointestinal stromal tumor. After post-operative imatinib treatment eosinophilia persisted, so that steroid therapy was started under an impression of hypereosinophilic syndrome. In spite of 6 months steroid therapy, eosinophilia persisted. Stool examination was performed to rule out intestinal helminth infections. Rhabditoid larvae of Strongyloides stercoralis were detected and the patient was diagnosed as strongyloidiasis. This diagnosis was confirmed again by PCR. The patient was treated with albendazole for 14 days and her abdominal pain and diarrhea improved. This case highlights the need for thorough investigation, including molecular approaches, to test for strongyloidiasis before and during steroid therapies.

Alteration of the SETBP1 Gene and Splicing Pathway Genes SF3B1, U2AF1, and SRSF2 in Childhood Acute Myeloid Leukemia

BACKGROUND: Recurrent somatic SET-binding protein 1 (SETBP1) and splicing pathway gene mutations have recently been found in atypical chronic myeloid leukemia and other hematologic malignancies. These mutations have been comprehensively analyzed in adult AML, but not in childhood AML. We investigated possible alteration of the SETBP1, splicing factor 3B subunit 1 (SF3B1), U2 small nuclear RNA auxiliary factor 1 (U2AF1), and serine/arginine-rich splicing factor 2 (SRSF2) genes in childhood AML. METHODS: Cytogenetic and molecular analyses were performed to reveal chromosomal and genetic alterations. Sequence alterations in the SETBP1, SF3B1, U2AF1, and SRSF2 genes were examined by using direct sequencing in a cohort of 53 childhood AML patients. RESULTS: Childhood AML patients did not harbor any recurrent SETBP1 gene mutations, although our study did identify a synonymous mutation in one patient. None of the previously reported aberrations in the mutational hotspot of SF3B1, U2AF1, and SRSF2 were identified in any of the 53 patients. CONCLUSIONS: Alterations of the SETBP1 gene or SF3B1, U2AF1, and SRSF2 genes are not common genetic events in childhood AML, implying that the mutations are unlikely to exert a driver effect in myeloid leukemogenesis during childhood.

Constitutional Chromosomal Abnormality Identified in a Sibling Donor After Bone Marrow Stem Cell Transplantation in a Pediatric Patient with Acute Megakaryoblastic Leukemia

No abstract available.

Molecular Identification and Amphotericin B Susceptibility Testing of Clinical Isolates of Aspergillus From 11 Hospitals in Korea

BACKGROUND: We investigated the species distribution and amphotericin B (AMB) susceptibility of Korean clinical Aspergillus isolates by using two Etests and the CLSI broth microdilution method. METHODS: A total of 136 Aspergillus isolates obtained from 11 university hospitals were identified by sequencing the internal transcribed spacer (ITS) and beta-tubulin genomic regions. Minimal inhibitory concentrations (MICs) of AMB were determined in Etests using Mueller-Hinton agar (Etest-MH) and RPMI agar (Etest-RPG), and categorical agreement with the CLSI method was assessed by using epidemiological cutoff values. RESULTS: ITS sequencing identified the following six Aspergillus species complexes: Aspergillus fumigatus (42.6% of the isolates), A. niger (23.5%), A. flavus (17.6%), A. terreus (11.0%), A. versicolor (4.4%), and A. ustus (0.7%). Cryptic species identifiable by beta-tubulin sequencing accounted for 25.7% (35/136) of the isolates. Of all 136 isolates, 36 (26.5%) had AMB MICs of > or =2 microg/mL by the CLSI method. The categorical agreement of Etest-RPG with the CLSI method was 98% for the A. fumigatus, A. niger, and A. versicolor complexes, 87% for the A. terreus complex, and 37.5% for the A. flavus complex. That of Etest-MH was < or =75% for the A. niger, A. flavus, A. terreus, and A. versicolor complexes but was higher for the A. fumigatus complex (98.3%). CONCLUSIONS: Aspergillus species other than A. fumigatus constitute about 60% of clinical Aspergillus isolates, and reduced AMB susceptibility is common among clinical isolates of Aspergillus in Korea. Molecular identification and AMB susceptibility testing by Etest-RPG may be useful for characterizing Aspergillus isolates of clinical relevance.

Factors Predicting Peripheral Blood Stem Cell Collection: Analysis of Korean Patients at a Single Center / 대한수혈학회지

BACKGROUND: Peripheral blood stem cell (PBSC) transplantation is a curative treatment in various hematologic malignancies and some solid cancers. Effective mobilization and collection of PBSC is essential for successful PBSC transplantation. The aim of this study was to investigate the useful factors for predicting PBSC collection using multivariate analysis. METHODS: We retrospectively reviewed the medical records of 170 allogeneic and 389 autologous donors at Chonnam National University Hwasun Hospital between 2005 and 2012. Donor groups were divided into three groups (failure group, suboptimal group, and optimal group) according to the total CD34+ yield. Donors were compared regarding age, sex, body weight, disease, complete blood count, hematopoietic progenitor cell (HPC) parameter of automated cell counter, process volume, number of leukapheresis procedures, prior mobilization history, type of vascular access and instrument. RESULTS: In allogeneic PBSC collections (n=170), the collection failure group showed lower baseline (premobilization) white blood cell (WBC) (P=0.004) and HPC (P<0.001) than the optimal group. In autologous PBSC collections (n=389), the collection failure group showed lower baseline HPC and more frequent prior mobilization history (P<0.001) than the suboptimal and optimal group. In multivariate analysis, older age, lower number of leukapheresis procedures, and prior mobilization history were risk factors associated with mobilization failure. CONCLUSION: Our data suggest that baseline WBC and HPC would be useful for predicting poor mobilizer in allogeneic PBSC collection, whereas baseline HPC would be useful in autologous PBSC collection. Conventional chemotherapy and G-CSF based remobilization would not be helpful to proven poor mobilizer in previous mobilization.

The First Known Case of Blood Group Chimerism in Monochorionic Dizygotic Twins in Korea

No abstract available.

Direct Measurement of Serum Immunoglobulin Heavy and Light Chain Pairs for Identification of Monoclonal Gammopathy and a Performance Comparison with Capillary Electrophoresis

BACKGROUND: Determination of monoclonal gammopathy through conventional protein electrophoresis is sometimes difficult because of the presence of large proteins such as haptoglobin and transferrin, which may obscure the results. Ambiguity in an electrophoresis band can give rise to confusion or difficulty in interpretation. The heavy chain/light chain assay (HLC assay) using Hevylite antibody (The Binding Site, UK) has recently been developed for the accurate measurement of monoclonal proteins. We compared the immunotyping (IT) profiles to the immunoglobulin (Ig) heavy/light chain measurements obtained using the HLC assay and observed the ratios between intact Ig kappa and lambda. METHODS: We collected 35 and 28 sera from patients with suspicious and definitive monoclonal protein, respectively. Then we performed serum protein electrophoresis (SPEP) and IT by Capillarys2 (Sebia, USA). Monoclonal protein production was investigated using Freelite antibody (The Binding Site) and specific Ig(G, A)kappa and Ig(G, A)lambda Hevylite antibodies. The results were analyzed using PASW 18.0 for Windows (IBM, USA). RESULTS: Direct measurement of Ig heavy/light chains showed discordant IT results for 12 (34.2%) of 35 patients' sera with suspicious SPEP pattern and identical IT results for 28 patients' sera with definitive monoclonal peak in the SPEP results. Overall, the results of the HLC assay and IT showed good agreement (kappa=0.718, P=0.000 by cross-tabulation Gamma, Kappa analysis). CONCLUSIONS: The results of direct measurement of serum Ig heavy chain/light chain pairs were comparable to those of IT and were helpful for determination of monoclonality in the case of ambiguous electrophoresis results. Measurement of the heavy chain/light chain pair ratio also allowed precise quantification of the monoclonal Igs with ambiguous electrophoresis patterns and identification or discrimination of clonality.

Congenital Chimera and Blood Group: An Analysis of the Korean Cases in the Literature / 대한수혈학회지

No abstract available.

Transfusion Support of Patients with Antibodies Against High-Frequency RBC Antigens / 대한수혈학회지

No abstract available.

Autoantibodies with Mimicking Specificity Detected by the Dilution Technique in Patients with Warm Autoantibodies

BACKGROUND: The aim of this study was to investigate the frequency of autoantibodies with mimicking specificity by using the dilution technique, to assess the usefulness of the combination of the dilution technique and red blood cell (RBC) phenotyping, and to establish a pre-transfusion testing algorithm in patients with warm autoantibodies. METHODS: Serum samples from 71 patients with warm autoantibodies were tested using the dilution technique. Among them, 25 samples were adsorbed with allogeneic ZZAP (a combination of dithiothreitol and enzyme) or polyethylene glycol (PEG) and their RBC phenotypes were determined. Thirty-nine patients were transfused with our pre-transfusion testing algorithm using a combination of dilution technique and RBC phenotyping. RESULTS: Autoantibodies with mimicking specificity were detected by the dilution technique in 26.8% (19/71) of the patients and most of them were directed against Rh system antigens. The agreement of the results obtained with the dilution technique in combination with RBC phenotyping and those from ZZAP or PEG adsorption was 100% (18/18) in patients who have autoantibodies with mimicking specificity and/or alloantibodies. No clinical symptoms indicating severe acute or delayed hemolytic transfusion reactions were reported in the 39 patients transfused with our pre-transfusion testing algorithm. CONCLUSIONS: Autoantibodies with mimicking specificity detected by the dilution technique in patients with warm autoantibodies are relatively frequent, can be discriminated from alloantibodies by employing a combination of dilution technique and RBC phenotyping, and might not appear to cause severe acute or delayed hemolytic transfusion reactions.

Analysis of ABO Blood Discrepancies and Transfusion Experiences in Chonnam National University Hospital / 대한수혈학회지

BACKGROUND: ABO blood group discrepancy occurs when the results of red cell tests do not agree with those of the serum test. In order to select the proper blood units for transfusion, clarification of the cause of ABO discrepancies is essential. We analyzed the cases and recent actual transfusion experiences at Chonnam National University Hospital (CNUH). METHODS: In total, among pre-transfusion blood samples at CNUH between January 2012 and July 2013, 55 cases of ABO discrepancies were analyzed retrospectively. RESULTS: The discrepancy incidence was 0.14%. Problems with serum were the most common cause of ABO discrepancies, with 31 cases (56.4%), and extra serum reactivity due to cold allo-antibodies accounted for the highest frequency (n=7). There were three cases of non-specific aggregations caused by commercial RBC constituents and aggregation was not observed when a re-test was performed with other commercial RBCs or self-prepared human RBCs. Two of three cases with mix-field aggregations involved a pair of twins after in vitro fertilization - embryo transfer (IVF-ET). Among 55 patients, 20 were actually transfused, and all but four cases had weaker or identical RBC units and stronger or identical plasma units. CONCLUSION: There were newly revealed ABO discrepancies caused by non-specific aggregations of commercial RBCs and in twins after IVF-ET. In addition, investigation of actual transfusion experiences in patients with ABO blood group discrepancies would be helpful.

A Case of ABO* cis-AB01/Ax03 Found in A2B3 Phenotype / 대한수혈학회지

The cis-AB blood group is rare; however, it is relatively more common in the Korean and Japanese populations. Among nine cis-AB alleles, only the cis-AB01 allele has been reported in the Korean population. When the A2B3 phenotype is found, it has the cis-AB01/O genotype; to date, it has not been reported in any other genotype. Here we report on an extremely rare case of cis-A2B3 found in the cis-AB01/Ax03 genotype. EDTA samples of a 52-year-old male with hepatocellular carcinoma and his family were sent to our laboratory. Standard ABO typing and sequencing of exons 6 and 7 of the ABO gene of the propositus and his family were performed: the propositus with the A2B3 type had cis-AB01/Ax03, brother with O type had O01/O02, sister with O type had Ax03/O01, son with B type had Ax03/B101, and daughter with A1B3 type had cis-AB01/A102. Results based on family study and genotyping revealed that the propositus had both cis-AB01 and Ax03 alleles. This is the first case of A2B3 phenotype with cis-AB01 with an allele other than the O allele in the cis-AB blood group reported so far.