Survey on Transfusion Laboratory Work inMedical Institutions Participating in the ExternalQuality Control Program for Blood Bank

Background@#Laboratory tests in blood banks vary with respect to methods,equipment, and quality control according to the hospital’s environment. @*Methods@#We surveyed institutions that regularly participated in the Koreanassociation of external quality assessment using a web-based questionnairecomprising 79 questions regarding transfusion laboratory work. @*Results@#A total of 84 institutions were surveyed including 17 senior generalhospitals, 43 general hospitals, 19 hospitals, four clinics, and one commerciallaboratory. ABO cell typing was performed by slide (63, 75.0%), tube (42,50.0%), automated column (19, 22.6%), and automated microplate (7, 8.3%)methods. ABO serum typing was performed by tube (75, 89.3%), automatedcolumn (19, 22.6%), automated microplate (7, 8.3%), and slide (7, 8.3%)methods. Irregular antibody screening test and identification test wasperformed by 58 (69.0%) and 36 (42.9%) institutions, respectively. Irregularantibody screening test and identification test was performed by the columnagglutination method in 34 (40.5%) and 26 (31.0%) institutions, respectively.Room temperature saline, albumin, and anti-globulin reagent crossmatchingtest (three-step method) was the most popular method (48, 57.1%). Theuse of anti-globulin reagent in the crossmatching test did not significantlyvary according to the size of the hospital. A daily quality control programfor ABO, Rh typing, and the crossmatching test was conducted in 58 (69.0%)institutions. @*Conclusions@#There were differences in transfusion-related laboratory testsamong the institutions. Although this survey included a limited number ofinstitutions, it can be helpful to evaluate the routine laboratory tests andtransfusion-related blood bank work in each institution.

Beta-Propeller-Protein-Associated Neurodegeneration: A Case of Mutation in WDR45

No abstract available.

Very Early Onset of Amiodarone-Induced Pulmonary Toxicity

Amiodarone is a widely used antiarrhythmic agent. Among its various adverse effects, amiodarone-induced pulmonary toxicity (APT) is the most life threatening complication, which has been described mostly in patients who have been in treatment with high accumulative doses for a long duration of time. However, amiodarone therapy in short-term duration induced APT was rarely reported. We describe a case of a 54-year-old man who is presented with symptoms of APT after a few days of therapy for post-myocardial infarction ventricular tachycardia. For early diagnosis and successful treatment, awareness and high suspicion of this rare type of early onset APT is crucial in patients with amiodarone therapy.

Diagnostic Role of C-reactive Protein, Procalcitonin and Lipopolysaccharide-Binding Protein in Discriminating Bacterial-Community Acquired Pneumonia from 2009 H1N1 Influenza A Infection / 결핵및호흡기질환

BACKGROUND: It is difficult but important to differentiate between bacterial and viral infections, especially for respiratory infections. Hence, there is an ongoing need for sensitive and specific markers of bacterial infections. We investigated novel biomarkers for discriminating community acquired bacterial pneumonia from 2009 H1N1 influenza A infections. METHODS: This was a prospective, observational study of patients with community acquired bacterial pneumonia, 2009 H1N1 Influenza A infection, and healthy controls. Serum samples were obtained on the initial visit to the hospital and stored at -80degrees C. We evaluated CRP (C-reactive protein), PCT (procalcitonin), LBP (lipopolysaccharide-binding protein) and copeptin. These analytes were all evaluated retrospectively except CRP. Receiver operating characteristic curve (ROC) analyses were performed on the resulting data. RESULTS: Enrolled patients included 27 with community acquired bacterial pneumonia, 20 with 2009 H1N1 Influenza A infection, and 26 who were healthy controls. In an ROC analysis for discriminating community acquired bacterial pneumonia from 2009 H1N1 influenza A infection, areas under the curve (AUCs) were 0.799 for CRP (95% Confidence interval [CI], 0.664~0.934), 0.753 for PCT (95% CI, 0.613~0.892) and 0.684 for LBP (95% CI, 0.531~0.837). Copeptin was not different among the three groups. CONCLUSION: These findings suggest that serum CRP, PCT and LBP can assist physicians in discriminating community acquired bacterial pneumonia from 2009 H1N1 influenza A infection.

Three Reinfection Cases of the Pandemic Influenza (H1N1 2009) / 감염과화학요법

Infection with influenza virus results in acquisition of immunity, preventing reinfection with the homologous virus. Although reinfection following primary infection is rare, its incidence depends on immunity of human body, antigenic diversity of influenza virus, and the presence of outbreak in the community. During the pandemic influenza (H1N1 2009), a child and two women were reinfected by H1N1 influenza virus several weeks after the primary infection, and they were successfully treated again by oseltamivir. This case series will provide additional information on diagnosis, treatment, and prevention of the pandemic influenza (H1N1 2009).

Down-Turner Syndrome (45,X/47,XY,+21): Case Report and Review / 대한진단검사의학회지

We report the case of a 3-yr-old boy with Down-Turner mosaicism and review the previous reports of Down-Turner syndrome with documented karyotyping and clinical features. The patient showed clinical features of Down syndrome without significant stigma of Turner syndrome. Cytogenetic analysis of peripheral blood preparations by using G-banding revealed mosaicism with 2 cell lines (45,X[29]/47,XY,+21[4]). FISH analysis revealed that 87.5% of the cells had monosomy X karyotype and 12.5% of the cells had XY karyotype; trisomy 21 was only detected in the Y-positive cells. We suggest that additional cells should be analyzed and molecular genetic studies should be conducted to rule out double aneuploidy when karyotypes with sex chromosome aneuploidies and mosaicism are encountered, as in our case of Down syndrome mosaic with sex chromosome aneuploidy.

Early Extracorporeal Membrane Oxygenation in a Patient with Pandemic Influenza (H1N1 2009) and Acute Respiratory Distress Syndrome / 감염과화학요법

Despite advanced technologies in intensive care, pandemic influenza (H1N1 2009) can rapidly progress to acute respiratory distress syndrome (ARDS) and cause death in a small subset of patients. Extracorporeal membrane oxygenation (ECMO) is expected to provide adequate gas exchange, to reduce ventilator-induced lung injury and, eventually, to improve outcome in these patients. A previously healthy, young female received mechanically ventilatory support because of rapidly progressive respiratory failure caused by 2009 H1N1 influenza. As she failed to respond to high ventilatory support, ECMO was instituted at 6 hours after admission. We describe detailed course of case and literature review on ECMO, helping physicians make a decision to initiate ECMO in patients with influenza-related ARDS.

A Case of Immune Thrombocytopenic Purpura Developing after Treatment for Non-Hodgkin's Lymphoma

Immune thrombocytopenic purpura (ITP) can be classified as primary or secondary according to the presence of an underlying non-malignant or malignant disorder, including lymphoproliferative disorders. The estimated prevalence of ITP in patients with Hodgkin's lymphoma is about 1%, and its clinical course has been reported in approximately 50 patients. ITP is an unusual and poorly documented complication in patients with non-Hodgkin's lymphoma. Some cases have been described in patients who have undergone high-dose chemotherapy and autologous bone marrow/peripheral blood stem cell transplantation. Rare cases appear to be coincidental. Here, we report on a rare case of a 61-year-old man who had ITP after being in a state of complete remission of non-Hodgkin's lymphoma for about 15 months.

Evaluation of the Genotypes of the Lewis Blood Group in a Korean Population Using Direct Sequencing

BACKGROUND: The FUT2 and FUT3 genes determine the Lewis phenotype of red blood cells (RBCs). Recently, the Lewis genes, the secretor genes, and several mutations that cause Lewis negative and nonsecretor phenotypes have been identified. The purpose of this study was to analyze the gene frequency of FUT2 and FUT3 in a Korean population by comparing the use of the direct sequencing method to the use of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for mutation detection in the FUT2 and FUT3 genes. METHODS: RBCs and peripheral blood leukocytes were obtained from 225 apparently healthy volunteers to determine the phenotype and genotype of the FUT2 and FUT3 genes. Lewis phenotypes were determined on K3EDTA-stablized fresh blood samples using the column agglutination method. Lewis blood group genotyping was performed by use of the direct sequencing method. For the detection of T59G, C357T, and A385T mutations, the PCR-RFLP method was performed. RESULTS: The frequencies of the Lewis blood group phenotype were 12.4% for Le(a+b-), 70.7% for Le(a-b+), 11.1% for Le(a-b-) and 5.8% for Le(a+b+), respectively. Direct Sequencing of the FUT2 gene identified 92.2% C357T, 56.9% A385T, 0.4% G244A mutations and 1.8% del396. Direct Sequencing of the FUT3 gene identified 46.9% T59G, 30.4% G508A, 1.1% T202C, 1.1% C314T, 0.7% A1029G, 3.0% T1067A and 13.3% G1242A mutations. The PCR-RFLP method results were discordant in nine cases (1 case for C357T, 4 cases for A385T and 2 cases for T59G) as compared to the direct sequencing method results. CONCLUSION: We have determined the frequencies of FUT2 and FUT3 gene mutations in a Korean population. The use of the direct sequencing method was more accurate than the use of the PCR-RFLP method for the determination of the genotype of the FUT2 and FUT3 genes.

Protein Profile Changes in Platelet Concentrates According to Storage and Leukoreduction- Analysis Using Proteomics Technology / 대한진단검사의학회지

BACKGROUND: Knowing how the protein profile of platelet products changes with storage or leukoreduction may give us greater insight into cell physiology and the cause of transfusion reactions other than cytokines and chemokines. METHODS: We filtered four packs of platelet concentrates (PC) within 24 hr of blood collection and after 120 hrs of storage. Four aliquots of each supernatant in PC were obtained: pre-storage+prefiltration, pre-storage+post-filtration, post-storage+pre-filtration and post-storage+post-filtration. Routine chemistry tests and a two-dimensional electrophoresis (2-DE) were performed. The stained images were analyzed and the significant spots were identified using a peptide mass finger printing (PMF) with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis after trypsin digestion. RESULTS: The protein spots increased with storage and decreased after filtration (P<0.05, prestorage+post-filtration). The spot density of various proteins, including macrophage inflammatory protein-2 alpha, megakaryocyte colony stimulating factor and interleukin-22 changed with storage and leukoreduction. CONCLUSIONS: The database of identified protein spots and their changes produced in this study is a useful basic tool for future studies on the mechanism of transfusion reactions. Further studies should validate the significance of each protein spot.

Development of a New Blood Typing Kit Using the Microfluidics Separation Technique

BACKGROUND: Blood typing is an essential test for transfusion. Generally, blood typing is performed using a slide test, tube test or microcolumn agglutination test. The aims of this study were to develop a new blood typing kit using micromachining, microfluidics and microseparation methods, and to evaluate the clinical usefulness of the new blood typing kit. METHODS: We designed and manufactured a blood typing microchip using polydimethylsiloxane (PDMS), which contained a microchannel (25~200 micrometer). The blood sample and antisera to be tested were dropped on the microwell for movement and mixing by capillary action. Once agglutination occurred, the microchannel acts as a filter and the blood type was determined by observation by the naked eye. To evaluate the newtyping kit, we tested sensitivity using artificially diluted blood and compared the results of the new typing method with the slide and tube methods using 70 samples. RESULTS: The new blood typing kit could differentiate a +4~+2 agglutination reaction, but could not detect a +1 agglutination reaction as observed by the naked eye. Among 70 samples, the results of ABO and Rh typing by the new typing method (n=66, > or = +2 agglutination reaction by the column agglutination method) were in accord with the results of the tube and slide methods, but couldnot detect agglutination in all 4 clinical samples, below a +1 agglutination reaction. CONCLUSION: The new blood typing kit is inadequate for routine use in the clinical laboratory due to low sensitivity, but with further improvement, it can be used economically, conveniently and objectively for blood typing without any special equipment. Moreover, the microfludics and separation method may be broadly applicable in other tests using the hemagglutination method.

New Leukocyte Counting Method of Cerebrospinal Fluid: Using Transparent Ruler Tape / 대한진단검사의학회지

BACKGROUND: To enumerate leukocyte count in cerebrospinal fluid (CSF) is important for diagnosing bacterial meningitis. Using automated hematology analyzer for enumeration of leukocyte in CSF is below the sensitivity, so microscopic hemocytometric method is standard method. But this requires sufficient practical experience and has limitation of accuracy and stability. So we developed new manual method and evaluated it. METHODS: We designed new method using transparent ruler tape. We performed correlation, accuracy and precision test by counting leukocyte in diluted EDTA blood with three methods: new method, Neubauer and Nageotte hemocytometry. Twenty two CSF were used for stability test, which determines leukocyte count according to time (within one hour and after 2, 4 and 12 hr), by new method and Neubauer hemocytometry at room temperature. RESULTS: There was no clinical significant difference between three methods in correlation test, whereas Neubauer and Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with new method in case with low leukocyte count. The new method showed the lowest CV and most accurate result. In stability test, leukocyte counts decreased being 44.4%, 72.1% of initial values after 2 hr, 14.8%, 31.1%, after 4 hr and 4.2%, 8.7%, after 12 hr, by Nageotte hemocytometry and new method, respectively. CONCLUSIONS: The new method we devised is simple, easy and applicable to use in a laboratory and offers advantages of improved precision and stability. It may be sufficient for replacing standard methods for leukocyte counting in CSF.

The Reduction Effect of Leukocyte Fragments by Leukodepletion Filter / 대한수혈학회지

BACKGROUND: The efficiency of leukocyte removal filter is influenced by many factors. But, filtration efficiency of leukocyte fragments was not well known. We performed this study to evaluate whether the filtration efficiency for packed red blood cells can be influenced by leukocyte fragments according to storage time. METHODS: Leukocyte fragments in packed red blood cells (three units) which were artificially made by incubation for 4 hrs at 56degrees C and each four units of packed red blood cells according to storage time (0 days, 10 days, 20 days, and 30 days) were filtered using Sepacell R-500A (Asahi medical Co, Japan). The leukocyte concentrations of the pre-leukodepleted samples were estimated using an automated hematology analyzer (XE-2100, Sysmex, Japan). The ratio between the number of normal leukocytes and leukocyte fragments on Wright Giemsa stained slide was used in the analysis. The leukocyte concentrations of the post-leukodepleted samples were performed by the conventional counting methods using Nageotte hemocytometer. RESULTS: The ratios of fragmented to total leukocytes in packed red blood cells at pre- and post leukoreduction according to storage times were 1.5% and 16.3% within 1 days, 4.5% and 30.0% at 10 days, 6.3% and 35.0% at 30 days, and 8.3% and 42.5% at 40 days, respectively. Leukoreduction efficiencies of normal leukocytes in packed red blood cells were 99.99 +/- 0.01%, 99.97 +/- 0.02%, 99.98 +/- 0.01%, and 99.86 +/- 0.09%, respectively. The 36.0% of leukocytes in packed red blood cells were changed to fragmented leukocytes, residual fragmented leukocytes ratio was 95.0% and filter efficiencies of normal leukocytes was low(99.28%, p<0.05). CONCLUSIONS: The leukodepleted efficiency for leukocyte fragments were lower than for normal leukocytes. Leukocytes fragments may be influenced to lower the leukodepleted efficiency of normal leukocytes with storage time elapse.

Two Cases of ABO Discrepancy due to Hypogammaglobulinemia / 대한수혈학회지

Two cases of ABO discrepancy were observed in thirty-year old woman with gall bladder abscess and fifty-five-year old woman with hepatocellular carcinoma. Their red cells were typed as group O and their serum had only anti-A antibody. Absence of A and B antigens on their RBCs were confirmed by adsorption elution test and saliva test. The B transferase activities were not demonstrated in their serum. Their ABO genotypes were O/O by sequence specific polymerase chain reaction. Their serum protein electrophoresis showed hypogammaglobulinemia pattern, and immunoglobulin levels (IgG, IgA, IgM) were decreased (39 mg/dL, 46 mg/dL, <5 mg/dL and 63 mg/dL, 65 mg/dL, 12 mg/dL, respectively).

A Case of Plasmodium vivax Infection Diagnosed after Treatment of Imported Falciparum Malaria / 대한임상병리학회지

In South Korea, most imported malaria has been caused by Plasmodium falciparum and little mixed infection has been reported which is frequently observed in other countries. We report a case of mixed malarial infection with P. falciparum/P. vivax. It was diagnosed by using three kinds of dipstick tests (two malarial antibody tests and one pLDH immunocapture assay), polymerase chain reaction (PCR), and a peripheral blood smear in a nineteen-year-old woman who had traveled to Bangladesh. The patient was diagnosed with P. falciparum on the first-visit blood sample and P. vivax with the second-visit sample by using all three kinds of dipstick tests. Peripheral blood smears and a PCR of the patient's blood also showed P. falciparum and P. vivax parasites and DNA bands of two species of malaria. In conclusion, an imported malaria case should be accurately diagnosed using the malarial antibody detection kit, pLDH immunocapture assay, and PCR in addition to the peripheral blood smear for the possibility of mixed infection with malaria.

A Case of Knee Joint Infection by Mycobacterium abscessus / 대한임상병리학회지

Rapid growing mycobacterium grows in less than 7 days on most types of solid media including the Ogawa media. Ninety percent of human diseases caused by rapid growing mycobacterium are due to Mycobacterium abscessus, Mycobacterium chelonae and Mycobacterium fortuitum. We report an isolated case of wound infection due to M. abscessus following total knee replacement arthroplasty surgery. The patient has presented arthralgia and fever for 3 weeks. From the joint fluid aspirates, pale gram-positive beaded rods were found but cultures were negative after 24 hours. After 48 hours, microorganisms grew on blood agar plates as tiny pinpoint colonies and they had odor of freshly-turned soil. They gave a positive reaction in a partial acid fast, an acid-fast stain and a heat-stable catalase but gave a negative reaction to PCR for IS6110. They were identified as the M. chelonae group biochemically and confirmed as M. abscessus through PCR-restriction fragment length polymorphism using restriction endonuclease, BstE II. Because rapid-growing mycobacterium can grow on a blood agar plate, an acid-fast stain should be selectively conducted in addition to a Gram stain in a microbiology laboratory.

Clinical Features and Molecular Epidemiology of Salmonella typhimurium for a Year Isolated in the Tertiary Hospital / 대한임상미생물학회지

BACKGROUND: The incidence of reported nontyphoidal Salmonellosis has increased during last decade in Korea. Salmonella typhimurium and Salmonella enteritidis are two major serotypes in nontyhoidal Salmonella. To determine the nature of potential outbreak S. typhimurium infection in a community, we retrospectively evaluated clinical and epidemiologic features of S. typhimurium infections and performed pulsed-field gel electrophoresis(PFGE) to investigate a genetic relatedness of S. typhimurium isolated in Guro Hospital. METHODS: From May 1998 to April 1999, a total of 20 S. typhimurium strains were isolated from 18 patients. PFGE patterns were analyzed for 20 S. typhimurium strains Clinical and epidemiological features were evaluated from their medical records. RESULTS: Seventy two percent(13/18) were acute gastroenteritis, and 11 %(2/18) were enteric fever and 16 %(3/18) were intussusception. Seventy eight percent(14 of 18) of patients were six years old or less than. There were two major type(A, B) on PFGE analysis. Eight of 20 strains showed identical PFGE type(A1). Eleven strains were subtypes of A1. One strain showed different type(B). Similarity coefficients between A1 and its subtypes were all over 0.765 and they showed close genetic distance on dendrogrm. Antibiogram of Al eight strains were various. CONCLUSIONS: High genetic relationship among 20 S. typhimurium strains for a year in Guro area indicates that they were possibly originated from one clone and that there might be a common source of infection. More efforts should be directed toward the epidemiological investigation of the cases to detect outbreaks and prevent further spread of the infection.