Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

BACKGROUND: Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are increasingly important in immunocompromised patients. Nucleic acid extraction methods could affect the results of viral nucleic acid amplification tests. We compared two automated nucleic acid extraction systems for detecting CMV and EBV using real-time PCR assays. METHODS: One hundred and fifty-three whole blood (WB) samples were tested for CMV detection, and 117 WB samples were tested for EBV detection. Viral nucleic acid was extracted in parallel by using QIAsymphony RGQ and QIAcube (Qiagen GmbH, Germany), and real-time PCR assays for CMV and EBV were performed with a Rotor-Gene Q real-time PCR cycler (Qiagen). Detection rates for CMV and EBV were compared, and agreements between the two systems were analyzed. RESULTS: The detection rate of CMV and EBV differed significantly between the QIAsymphony RGQ and QIAcube systems (CMV, 59.5% [91/153] vs 43.8% [67/153], P=0.0005; EBV, 59.0% [69/117] vs 42.7% [50/117], P=0.0008). The two systems showed moderate agreement for CMV and EBV detection (kappa=0.43 and 0.52, respectively). QIAsymphony RGQ showed a negligible correlation with QIAcube for quantitative EBV detection. QIAcube exhibited EBV PCR inhibition in 23.9% (28/117) of samples. CONCLUSIONS: Automated nucleic acid extraction systems have different performances and significantly affect the detection of viral pathogens. The QIAsymphony RGQ system appears to be superior to the QIAcube system for detecting CMV and EBV. A suitable sample preparation system should be considered for optimized nucleic acid amplification in clinical laboratories.

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

Diagnostic genetics subcommitee of KSQACP has performed two trials each for cytogenetic studies and molecular genetic studies in 2005. Cytogenetic surveys were performed by 37 laboratories and answered correctly in most laboratories. And the first trial with whole blood specimen from the patient with cytogenetic abnormality (mosaic Turner syndrome) was successfully done, which could evaluate the whole process for cytogenetic studies from cell culture, harvest and karyotyping.. The molecular genetic test surveys included many kinds of tests like M. tuberculosis, HBV, HCV, HPV, leukemias/lymphomas, ApoE genotyping, MTHFR genotyping, BRCA1 & BRCA2, Duchenne muscular dystrophy, and Huntington disease. Molecular genetic surveys showed excellent results in most of participants. External quality assessment program for genetic analysis in 2005 was proved to be helpful in continuous education and evaluation of quality improvement.

Annual Report on External Quality Assesment in Hematology in Korea (2004) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2004 with about 440 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 and coagulation test. The response rate was more than 96%. The coefficients of variation in hemoglobin and RBC number was stable but variable in platelet number and WBC number according to measuring cell counts. Blood coagulation study was performed twice. Test results show wide variation according to measuring machine and reagents.

Annual Report on External Quality Assesment in Hematology in Korea (2003) / 임상검사와정도관리

Four trials of external quality assessment in diagnostic hematology were performed in 2003 with about 430 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 and coagulation test. The response rate was more than 95%. The performance of quality assessment appeared to be gradually improved year by year.

Accelerated Phase Chronic Myeloid Leukemia in Complete Molecular Response with STI571 / 대한혈액학회지

STI571 is an effective agent for the patients with chronic myeloid leukemia (CML). But, complete molecular response with STI571 is rarely reported in accelerated phase CML. Here we report a patient with accelerated phase CML who achieved complete molecular response with STI571. A 60-year old female patient visited emergency room with syncope. Her white blood cell count was 30,800/microliter (basophil 23%), hemoglobin 8.9g/dL, and platelet counts 2,748,000/microliter. Bone marrow was hypercellular with increase in megakaryocyte and basophils (15%). She was diagnosed as an accelerated phase CML. Seven days after stopping hydroxyurea, we used STI571 in a daily oral dose of 600mg. Generalized edema and skin rash were observed 15 days after treatment (all grade 1) and were controlled well with conservative management. Complete hematologic and cytogenetic responses were achieved after 1 month and 3 months of therapy with STI571 respectively. Complete molecular response was simultaneously proven by conventional reverse transcriptase PCR and real-time PCR analysis. The patient still remained in complete hematologic, cytogenetic, and molecular responses for 24 months. Treatment with STI571 was well tolerated and rapid hematologic improvement was observed. This case shows STI571 can induce complete molecular response as well as cytogenetic response in accelerated phase CML.

Vitamin D and estrogen receptor gene polymorphism and their interaction associated with bone mineral density in Korean postmenopausal women / 대한내과학회지

BACKGROUND: Strong genetic components to the determination of bone mineral density (BMD) have been suggested by family and twin studies. However, association between gene polymorphism and BMD was not consistent in Korean as well as other ethnic groups. The goal of present study is to evaluate the relationship between vitamin D receptor (VDR) and/or estrogen receptor (ER) gene polymorphism and BMD after adjusting for suggested confounding factors and the possibility of VDR gene by ER gene interaction which could impact the bone mass of Korean postmenopausal women. METHODS: We determined the VDR and ER genotypes using a polymerase chain reaction based Bsm I restriction length fragment polymorphism (RFLP) and Pvu II and Xba I RFLP respectively, in a population based DNA sample of 132 Korean postmenopausal women aged 45 to 71. And then related the VDR and ER genotypes to BMD, bone related hormones, biochemical bone markers, and clinical characteristics in these women. Multiple regression analysis was used to predict variables contributing to BMD. Age, height, weight, years since menopause, VDR B genotype, and ER P and X genotypes were used as independent variables. RESULTS: There was no significant relationship of VDR or ER genotypes to lumbar or femoral neck BMD, hormones, and bone turnover markers. However, after controlling for potential confounding factors, a statistically significant ER X genotype effect on femoral neck BMD (p=0.038), but not on lumbar BMD was observed. Moreover, there was more significant effect on femoral neck BMD by an interaction of VDR B * ER X genotype (p=0.013) in multiple regression analysis. CONCLUSION: The ER X genotype was associated with femoral neck BMD in Korean postmenopausal women. This association was more significant with the VDR B genotype interaction.

A Case of Cis-AB without B Antigen Expression / 대한수혈학회지

The cis-AB bood type is a rare phenomenon in which both the A and B blood types are inherited from a single parent. The cis-AB persons are not homogeneous with respect to reactivity of their red cells to anti-A and anti-B reagents, and are split into three groups with on the basis of the strength and characteristics of the serologic reactions; these reactivities are A2B3, A1B3 and A2B. A 7-year-old Korean boy was evaluated for paternity because he was presumptively identified as blood group AweakB and known blood types of his father and mother were A. In the repeated ABO blood typing, the child was typed as group A2B3 with weak anti-B, cis-AB being suspected. Both of his mother and father were typed as group A1 in cell and serum typing. In the saliva test and adsorption and elution studies of the parents, B substance was not detected. According to ABO genotyping, the child, mother and father showed cis-AB/O, A1/O and cis-AB/A1, respectively. The paternity was confirmed, but the father had unusual expression of cis-AB genotype. This was the second case of A1/cis-AB with phenotype A1, not expressing B antigen.

A Case of Greig Cephalopolysyndactyly Syndrome

Greig cephalopolysyndactyly syndrome (GCPS) is a disorder characterized by postaxial polydactyly of the hand, broad or occasionally bifid thumbs, preaxial polydactyly of the feet, broad halluces, syndactyly of the fingers or toes, macrocephaly, frontal bossing, hypertelorism and a broad nasal bridge. Intelligence is usually normal, although borderline IQ has been reported. Advanced bone age, mild hydrocephalus, craniosynostosis and agenesis of the corpus callosum are occasionally associated abnormalities. We report here a 10-day-old male infant with GCPS. Birth Weight was 2,400kg and gestational age was 39 wks. He had a wide broad high forehead, hypertelorism, broad nose base and cryptorchidism. He had preaxial polysyndactyly due to duplication of the right thumb and left accessory thumb, duplication of both halluces and syndactyly of both toes and fingers. His brain MRI showed corpus callosum agenesis, mild hydrocephalus and small choroid plexus cyst. High resolution chromosomal analysis showed a de novo balanced translocation 46, XY, t (7;8) (p22;q24.1). We report the first GCPS case in Korea with brief literature.

Korean Nationwide Surveillance of Antimicrobial Resistance of Bacteria in 1998

Antimicrobial resistance surveillance can provide information needed for empirical therapy of antimicrobial agents and for control of resistance. To determine the trend of antimicrobial resistance in Korea, in vitro susceptibility data in 1998 were collected from 25 hospitals participating to a program of Korean Nationwide Surveillance of Antimicrobial Resistance (KONSAR). The data were analyzed based upon hospital location and bed capacity. The results showed that cefoxitin-resistant E. coli and K. pneumoniae and 3rd-generation cephalosporin-resistant K. pneumoniae were prevalent, that 3rd-generation cephalosporin-resistant E. cloacae, S. marcesens and A. baumannii had increased, and ampicillin-resistant S. enterica were not rare. Oxacillin-resistant S. aureus, penicillin-non-susceptible pneumococci and beta-lactamase-producing H. influenzae were prevalent even smaller hospitals surveyed, and an increase of imipenem-resistant P. aeruginosa and vancomycin-resistant E. faecium is a new obvious threat. In general, resistance rates to some old antimicrobial agents, i.e., E. coli to ampicillin and S. aureus to oxacillin were high and did not vary greatly between the different levels of hospitals, while the rates to some of the newer ones, i.e., P. aeruginosa to imipenem, was quite variable and depended on the hospitals, probably reflecting difference in selective pressure.

TnI, cTnT, and CK-MB in patients with chronic renal failure / 대한내과학회지

BACKGROUND: Nonspecific elevations of CK-MB, cTnT have been well known in patients with chronic renal failure(CRF) on maintenance hemodialysis. It has been suggested that recently developed cTnI seldom shows nonspecific elevations in these patients. Status of CRF patients can be divided into three groups: predialysis group, hemodialysis group and peritoneal dialysis group. Until now, most researchers have studied CK-MB, cTnT and cTnI only in CRF patients receiving maintenance hemodialysis. No previous studies have ever compared the differences of the nonspecific positivity of CK-MB, cTnT and cTnI according to the different status of CRF patients. METHODS: Nonspecific positive ratios of cTnI, cTnT, & CK-MB in were evaluated 20 predialysis patients, 13 CAPD patients and 20 hemodialysis patients. No one had had any evidence of myocardial ischemia during the previous 3 months before the study entry. The predialysis group was again divided into two groups according to the cut off level of serum creatinine of 3.0 mg/dl. Authors also compared the nonspecific positive ratios of cTnI, cTnT, CK-MB between diabetic CRF group and non diabetic CRF group. The sensitivity, specificity and false positive ratios of each enzymes were examined on and 6 hours after arrival in 21 CRF patients who visited the emergency room with the complaint of chest pain. RESULTS: 1) There were no nonspecific significant elevations of cTnI in CRF patients regardless of the status of CRF. But there were significant nonspecific elevations of CK-MB, cTnT in them. It was more marked in cTnT especially with the cut-off value of 0.1 ng/ml. 2) Nonspecific positive ratios of cTnT was significantly increased in diabetic CRF patients. 3) The sensitivity and specificity of cTnI were 100% and 93.3% each, which were significantly higher than those of CK-MB(83.3%, 66.7%) & cTnT(66.7%, 53.3%). CONCLUSION: In CRF patients, the nonspecific positive ratios of CK-MB, cTnT were higher than that of cTnI, and only cTnI did show significant specific elevations in all the CRF patients with acute myocardial infarction. It is likely that the status of CRF patients, dialysis mode, the sampling time point would not give significant changes in the nonspecific positive ratios of CK-MB, cTnT and cTnI.

Unusual Antigen Expression of Acute Leukemia / 대한혈액학회지

BACKGROUND: The recent advances in flow cytometric technology and the development of monoclonal antibodies have led to the important insights into the cell lineage and maturation stage of leukemia. The increased use of immunophenotyping in acute leukemia revealed the unusual anigen expression and biclonal or biphenotypic acute mixed lineage leukemia (AMLL). However, the data on their frequency and prognostic significance are still conflicting. METHODS: The immunophenotyping of leukemic cells (HLA-DR, CD10, CD19, CD20, CD22, CD3, CD5, CD7, CD13, CD33, CD61, TdT, cytoplasmic Ig, surface Ig) was performed by flow cytometry in 115 cases of acute leukemia between January 1994 and August 1996. Double-color immunofluorescent staining was performed in the cases expressing unusual antigens. RESULTS: 51 cases (44.3%) of 115 acute leukemias showed unusual antigens expression. These included 27 cases (38.6%) of 70 AML, 13 cases (43.3%) of 30 B-lineage ALL, 4 cases (50%) of 8 T-LL and 7 AMLL cases (6.1%) of 115 acute leukemias. CD7 (28.6%) and CD19 (11.4%) are expressed in AML, and CD13 (36.7%) and CD33 (26.7%) are expressed in ALL. Among 7 cases of AMLL, we could obtain the clinical data of 5 cases. The 4 cases of 5 AMLL failed to respond to induction chemotherapy or died before or during induction chemotherapy, and only one case showed partial remission. CONCLUSION: The unusual antigen expressions of acute leukemic cells are frequently observed, and the identification of relatively rare AMLL is very important, because AMLL showed poor response to the chemotherapy.

Clinical Utility of Adenosine Deaminase Isoenzyme Determinations in Leukemia / 대한임상병리학회지

BACKGROUND: Adenosine deaminase (ADA), an enzyme involved in purine metabolism, catalyzes the hydrolytic deamination of adenosine or 2-deoxyadenosine to inosine or 2-deoxyinosine. Human ADA consists of three molecular forms: ADA1, ADA1+CP, and ADA2. The two ADA isoenzymes represent two different gene products and have different tissue distributions, and their concentrations in serum appear to reflect different pathological conditions or physiological responses. Elevation of serum ADA activity has been described especially in leukemia and lymphoma. The purpose of this study was to evaluate the clinical utility of ADA isoenzyme determination in the diagnosis of leukemia. METHODS: We studied the activity of serum ADA and its isoenzyme in 44 leukemic patients. The study population consisted of 17 patients with acute lymphoblastic leukemia (ALL), 23 with acute myeloid leukemia (AML), and 4 with chronic myelogenous leukemia (CML). ADA isoenzyme was measured by erythro-9- (2-hydroxy-3-nonyl) adenine [EHNA] inhibitory assay using the Hitachi 7170 autoanalyzer. RESULTS: The rates of abnormally high total ADA activity were 100% for ALL, 60.8% for AML, and 50% for CML. In isoenzyme pattern, there was a clear difference between ALL and AML. High level of ADA1 activity was found in patients with ALL (P <0.01). The ADA1/ADA2 ratio was significantly higher (P <0.001) in ALL than AML. There was a correlation between ADA1 and absolute number of peripheral blasts in AML (r=0.840). CONCLUSIONS: It is concluded that the measurement of ADA isoenzyme may be a useful biochemical marker for leukemic diagnosis.

A Case of Wolf-Hirschhorn Syndrome with del (4) (p15.2) / 대한임상병리학회지

Wolf-Hirschhorn syndrome (WHS) is a congenital anomaly associated with partial deletion of distal short arm of chromosome 4, characterized by distinct facial anomalies ("Greek-Helmet"), mental and growth retardation and midline defects. The deletion of 2.2Mb area of 4p16.3 is thought to be the critical for WHS. It is reported that the sizes of 4p deletions are variable and the clinical findings are different from classic WHS according to the deleted portion. Recently proximal interstitial deletions have been infrequently reported. On fetal ultrasonography, a fetus of this case was found to have intrauterine growth retardation and multiple somatic anomalies including cleft palate, cleft lip, club foot, and hypoplastic thoracic cage with cardiomegaly, hypoplasia of right kidney and small stomach. At 36th week of gestation, the female baby was born by normal vaginal delivery, and her body weight was only 2.1kg. In spite of vigorous resuscitative measures, she died of respiratory failure on the second day of life. Karyotype was 46,XX,del (4) (p15.2). It is supposed that the size of deleted 4p in patients with WHS doesn't have substantial influence on the phenotype of the disease if the critical region in 4p16.3 is also deleted.

Simple ABO genotyping method using three polymorphic sites at the ABO locus / 대한수혈학회지

BACKGROUND: Recent progress in the molecular biology of the ABO blood group system has recognized the molecular basis of the red cell antigens and has provided a genetic model for ABO polymorphism at the molecular level. Genotyping methods with the basis of this genetic model were tested for korean blood donors. METHODS: Simple genotyping of the ABO blood group was performed in 253 healthy Korean blood donors by polymerase chain reaction-restriction enzyme length polymorphism (PCR-RFLP) and allele-specific PCR analysis of the only three polymorphic nucleotide positions (nps) 261, 526 and 803 of the ABO blood group gene. Two differen regions of the ABO gene, each of which contained a different polymorphic site (np 261 or 526) were amplified. Amplified products were digested with four restriction enzymes, two complementary pairs, Kpn I and Bst PI, and Bss HII and Ban I to analyze the np 261 and 526, respectively. The np 803 was analyzed by PCR with allele-specific primers. The PCR determined genotypes were compared with serologically determined phenotypes. RESULTS: The results were consistent for all individuals and not different from the proposed genetic model. The ABO genotypes of 253 healthy Korean blood donors were13 AA, 64 AO, 11 BB, 57 BO, 81 OO and 27 AB. CONCLUSIONS: The simple method used in this study may serve as a helpful tool for solving difficulties in serological ABO blood typing in the transfusion and forensic medicine, especially for cases with ABO discrepancy due to genetic variations. This method would be more efficient and informative for genotyping of the Korean population.

Korean Nationwide Surveillance of Antimicrobial Resistance of bacteria in 1997

Antimicrobial-resistant bacteria are known to be prevalent in tertiary-care hospitals in Korea. Twenty hospitals participated to this surveillance to determine the nationwide prevalence of resistance bacteria in 1997. Seven per cent and 26% of Escherichia coli and Klebsiella pneumoniae were resistant to 3rd-generation cephalosporin. Increased resistance rates, 19% of Acinetobacter baumannii to ampicillin/sulbactam, and 17% of Pseudomonas aeruginoa to imipenem, were noted. The resistance rate to fluoroquinolone rose to 24% in E. coli, 56% in A. baumannii and 42% in P. aeruginosa. Mean resistance rates were similar in all hospital groups: about 17% of P. aeruginosa to imipenem, 50% of Haemophilus influenzae to ampicillin, 70% of Staphylococcus aureus to methicillin, and 70% of pneumococci to penicillin. In conclusion, nosocomial pathogens and problem resistant organisms are prevalent in smaller hospitals too, indicating nosocomial spread is a significant cause of the increasing prevalence of resistant bacteria in Korea.

Numb Chin Syndrome in Malignant Disease / 대한혈액학회지

The numb chin syndrome (NCS) is characterized by chin or lower lip numbness restricted to the distribution of mental nerve (the distal trigeminal nerve). This uncommon neuropathy may be associated with neoplastic disease and usually appeared as a late manifestation of systemic malignancy, and it is an important sign for early diagnosis and prediction of clinical course and prognosis of hematologic malignancy. The numb chin syndrome is usually associated with a poor prognosis although various therapeutic strategies led to resolution of this syndrome. We report 2 cases of numb chin syndrome ; one in acute leukemia in early course of disease before diagnosis of leukemia and the other in leptomeningeal seeding of malignant lymphoma. The therapeutic response and prognosis were poor, a patient of malignant lymphoma expired in two months and a patient of acute leukemia is alive at present but the disease was relapsed in 5 months after complete remission.

The Cytogenetic Study of Acute and Chronic Leukemic Patients in Korea / 대한임상병리학회지

BACKGROUND: Because specific chromosomal abnormalities are associated with certain hematologic disorders, cytogenetic studies can help classifing the diseases, providing the clues of disease progression and being used to monitor remission after chemotherapy. In this study, cytogenetic analysis was performed. In acute and chronic leukemic patients in Korea and the results were compared with foreign cytogenetic reports, and the typical acute lymphocytic leukemia (ALL) and acute myelocytic leukemia (AML) associated chromosome aberrations were analysed by some calculated parameters to clarify if the specific chromosomal abberations in the specific types or subtypes of leukemias had diagnostic value or not. METHOD: Chromosome studies were done in bone marrow or peripheral blood samples by high resolution banding technique. Sensitivity, specificity, and positive and negative predictive values of finding or not finding a given aberration were calculated for followings : for the differential diagnosis between ALL and AML when a patient is known to have acute leukemia, for the differential diagnosis among AML and ALL FAB subtypes in a patient with known AML and ALL. RESULTS: The high positive predictive values (1.0) in the AML versus ALL comparison were found for -7, del(7) (q11-34q22-36), +8s, t(8;21) (q22;q22), t(15;17) (q22;q11), inv (16) (q13;q22) and -Y. Among the AML subtypes, the highest sensitivity, positive and negative predictive values were 0.85, 0.97, 0.94 for t(15;17) (q22;q11) in M3, respectively. The high positive predictive values and specificity in the ALL versus AML comparison were found for t(1;19) (q23;p13) ,t(4;11) (q21 ;23) and t(8; 14) (q24;q32) Among the ALL subtypes, the highest negative predictive value was 0.99 for t (8;14) (q24;q32) in L3. Among 398 CML cases, Philadelphia chromosome positive CML were shown in 81.9% that were classic t(9;22) (q34;all) (94.5%), complex variant traslocation(1.8%) and additional secondary chromosome aberrations (3.7%) . CONCLUSION: Total chromosomal aberration rate in acute and chronic leukemia in Korea was lower than that in foreign reports, but the patterns of chromosome aberrations were similar except for t(15;17) (q22;q11) in AML patients. Quantitativly calculated data of sensitivity, specificity and positive and negative predictive values in the specific chromosomal aberration might be used for diagnostic markers of acute leukemia.

Gene rearrangement of specific exons of immunoglobulin heavy chain joining region in B- lymphoid leukemias / 대한혈액학회지

No abstract available.

Whole blood cyclosporine measurement by fluorescence polarization immunoassay / 대한이식학회지

No abstract available.

A case of acute toxoplasmosis showing toxoplasma on peripheral blood smear / 대한혈액학회지

No abstract available.