BACKGROUND: The precore mutant hepatits B virus (HBV) cannot produce HBeAg due to the formation of transnational stop codon at nucleotide 1896 of the HBV-DNA precore region. This mutant has been detected worldwide in acute fulminant hepatitis, carrier and chronic HBV infections. It has been controversial whether the emergence of precore mutant HBV is related to the severity of the chronic hepatitis B or not. METHODS: To determine the prevalence and clinical implication of precore mutant infection, 137 HBsAg (+) patients including 12 acute hepatitis, 59 carriers, 41 chronic hepatitis, 15 liver cirrhosis, and 10 hepatomas were examined with mutation site specific assay-polymerase chain reaction (MSSA-PCR). The specificity for the detection of mutant by MSSA-PCR method was confirmed by direct sequencing of PCR products. RESULTS: The precore mutant HBV was detected in 67 of 137 (49%) subjects : none of 12 (0%) acute hepatitis patients, 17 of 59 (29%) carriers, 31 of 41 (76%) chronic hepatitis patients, 12 of IS (80%) liver cirrhosis patients, and 6 of 10 (60%) hepatoma patients. According to the status of serum HBeAg, the emergence rate of precore mutant HBV in HBeAg(-) cases was relatively higher than in HBeAg(+) cases with blood donor and chronic hepatitis. In anti-HBe (+) patients with chronic hepatitis, the precore mutant HBV was found regardless of ALT level in all patients. Emergence rate of precore mutant HBV was abruptly increased after 30 years of age. Among HBV-DNA (-) sera by DNA probe method, the core region of HBV was amplified in 94% of sera by MSSA-PCR method. CONCLUSIONS: The presence of precore mutant HBV may be related to the duration of HBV infections and there seems to be no causal relationship between the presence of precore mutant HBV and the severity of chronic hepatitis.
Blood Donors ; Carcinoma, Hepatocellular ; Codon, Terminator ; DNA ; Hepatitis ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens ; Hepatitis B, Chronic ; Hepatitis, Chronic ; Herpesvirus 1, Cercopithecine ; Humans ; Liver Cirrhosis ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity

Human parvovirus B19 is a single-strand DNA virus which causes erythema infectlosum, arthralgia, aplastic crisis in patients with red cell defect, chronic anemia in immunocompromised patients, and fetal hydrops in pregnant women . A 16-year-old women was referred to our hospital with pancytopenia and splenomegaly. In peripheral blood, spherocytosis and reitculocytopenia were observed. Many giant pronormoblasts with prominent inclusion bodies and deeply blue cytoplasm were observed but late erythroblasts were not observed in bone marrow smear. Osmotic fragility of patient's red cells was significantly increased. Human parvovirus Bl9 DNA was detected by polymerase chain reaction. Only with supportive therapy, pancytopenia was spontaneously resolved.
Adolescent ; Anemia ; Arthralgia ; Bone Marrow ; Cytoplasm ; DNA ; DNA Viruses ; Erythema ; Erythroblasts ; Female ; Humans* ; Hydrops Fetalis ; Immunocompromised Host ; Inclusion Bodies ; Osmotic Fragility ; Pancytopenia ; Parvovirus B19, Human ; Parvovirus* ; Polymerase Chain Reaction ; Pregnant Women ; Splenomegaly

No abstract available.
Anemia, Megaloblastic* ; Megaloblasts* ; Primary Myelofibrosis*

BACKGROUND: Heparin salts induce negative proportional bias according to anticoagulant concentration for analysis of ionized calcium (iCa) However, D-phenylalanyl -L-prolyl- L-arginine chloromethyl ketone (PPACK), a selective thrombin inhibitor, do not bind to ionized calcium. Therefore, we evaluated PPACK as an alternative anticoagulant to lithium heparin (Li-Hep) for analysis of ira, blood gases and electrolytes. METHODS: The concentration of iCa in whole blood anticoagulated with heparin was compared with that in serum of patients admitted to Chungbuk National University Hospital (n=27). The blood gases, electrolytes and iCa according to each anticoagulant concentration (Ll-Hep or PFACK) were analyzed. The concentrations of anticoagulated whole blood (Li-Hep; 50 kIU/L, PPACK ; 75 mumol/L) were compared with those of nonanticoagulated whole blood for blood gases, electrolytes and iCa (n=17), RESULTS: The results were as follows; whole blood anticoagulated with Li-Hep demonstrated -0.28+/-0.15 mmol/L (26.6%) bias for ira compared with serum. No bias according to each anticoagulated concentrations were observed in analysis of blood gases, potassium and chloride. Negative proportional bias for iCa and sodium in serum anticoagulated with Li-HeP was observed. In comparison, no bias for ira and sodium was observed with PPACK. No bias was observed in analysis of blood gas or electrolytes with each anticoagulated whole blood except for sodium and chloride that had clinically nonsignificant bias. Whole blood anticoagulated with Li-Hep demonstrated a consistent -0.08+/-0.02 mmol/L (6.3%) bias for ira compared with nonanticoagulated whole blood. In comparison, no bias was observed with PPACK-anticoagulated whole blood for iCa. CONCLUSIONS: We concluded that PPACK is an ideal anticoagulant without bias for analysis of iCa, blood gases and electrolytes.
Arginine ; Bias (Epidemiology) ; Calcium* ; Chungcheongbuk-do ; Electrolytes* ; Gases ; Heparin* ; Humans ; Lithium ; Potassium ; Salts* ; Sodium ; Thrombin

PURPOSE: The purpose of this study was to determine the frequency of CD4+CD25+FoxP3+ regulatory T cells (Treg) in the peripheral blood of patients with childhood chronic immune thrombocytopenic purpura (ITP) exhibiting thrombocytopenia and spontaneous remission. The findings of this study indicate the possibility of predicting spontaneous recovery and pathogenesis of childhood chronic ITP. METHODS: Eleven children with chronic ITP (seven thrombocytopenic and four spontaneous remission cases; mean age, 8.8 years; range, 1.7-14.9 years) were enrolled in this study. Five healthy children and eight healthy adults were included as controls. The frequency of Treg was evaluated by flow cytometry in the peripheral blood. RESULTS: In this study, four patients (36%) achieved spontaneous remission within 2.8 years (mean year; range, 1.0-4.4 years). The frequency of Treg was significantly lower in patients with persisting thrombocytopenia (0.13%+/-0.09%, P<0.05), than that in the patients with spontaneous remission (0.30%+/-0.02%), healthy adults controls (0.55%+/-0.44%), and healthy children controls (0.46%+/-0.26%). A significantly positive correlation was found between the frequency of Treg and the platelet count in children. CONCLUSION: These data suggest that a lower frequency of Treg contributes to the breakdown of self-tolerance, and may form the basis for future development of specific immunomodulatory therapies. Furthermore, Treg frequency has prognostic implication toward the natural course and long-term outcomes of childhood chronic ITP.
Adult ; Child ; Flow Cytometry ; Humans ; Immunomodulation ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic* ; Remission, Spontaneous ; T-Lymphocytes, Regulatory* ; Thrombocytopenia

No abstract available.
Eosinophilia*

The bacterial protein streptokinase binds to LD-M subunits, which shares a small region of homology with the site on plasminogen to which streptokinase is known to bind. We found an extra band of LD activity in CSF in a patient, suffering from meningitis due to Streptococcus pneumoniae. We performed a LD isoenzyme electrophoresis of the serum mixed with supernates from cultured broth of several species of streptococci. To investigate the effect on serum LD activity, we analyzed LD activity after the mixing of the serum with products of S. pneumoniae. S. pneumoniae, groups A and C beta hemolytic streptococci, revealed the extra band of LD activity at the origin site. The supernates of cultured broth of S. pneumoniae inhibited LD activity of the serum. Streptokinase or streptokinase-like substances can form complexes with LD in vivo after streptococcal infection, with consequent alteration of the LD isoenzyme pattern.
Bacterial Proteins ; Electrophoresis ; Humans ; L-Lactate Dehydrogenase* ; Meningitis* ; Plasminogen ; Pneumonia ; Streptococcal Infections ; Streptococcus pneumoniae* ; Streptokinase

BACKGROUND: This study compared three non-molecular methods for the detection of methicillin-resistance directly from blood cultures containing Staphylococcus aureus: penicillin-binding protein (PBP) 2a latex agglutination (LA), PBP2a immunochromatographic assay (ICA) and MRSA chromogenic medium (CM). METHODS: Fifty methicillin-resistant S. aureus (MRSA) and 50 methicillin-susceptible S. aureus (MSSA) were seeded into blood-culture bottles. When isolates returned a positive signal, 5 mL of culture was added to serum separator tubes and centrifuged at 1,300 g for 10 min. The pellets were then used as the inoculum for the PBP2a LA, MRSA-CM and PBP2a ICA. The pure colony was used for PBP2a LA test, additionally. RESULTS: The respective sensitivities and specificities were 98 and 100% for PBP2a ICA, and 100 and 100% for MRSA-CM in direct detection of MRSA from positive blood culture. The results of PBP2a LA test using pure colony were entirely compatible with those by mecA gene PCR but the PBP2a LA test using the pellets directly isolated from positive blood culture showed sometimes ambiguous agglutination; its sensitivity and specificity were 78 and 100%, if ambiguous results were scored as negative, and were 90 and 92%, if ambiguous results were scored as positive, respectively. CONCLUSION: For direct detection of MRSA in positive blood culture, MRSA-CM and PBP2a ICA provided excellent results. The PBP2a LA test using pure colony also gave excellent results but the PBP2a LA test by the direct method using pellet of positive blood culture was slightly less sensitive than the other two methods.
Adenosine ; Agglutination ; Immunochromatography ; Latex ; Methicillin Resistance ; Methicillin-Resistant Staphylococcus aureus ; Penicillin-Binding Proteins ; Polymerase Chain Reaction ; Seeds ; Sensitivity and Specificity ; Staphylococcus

BACKGROUND: Because extended-spectrum -lactamase (ESBL) producing strains can frequent-ly cause therapeutic failure and infectious outbreaks in hospitals, rapid and accurate detection of these strains are important. We compared the Vitek ESBL test with the NCCLS ESBL phenotypic confirmatory test by disk diffusion (NCCLS ESBL test) and double disk synergy test (DDST). METHODS: For a total of 316 clinical isolates composed of Escherichia coli (184), Klebsiella pneu-moniae (120) and Klebsiella oxytoca (12), we performed the Vitek ESBL test and the NCCLS ESBL test. For sixty-eight ESBL producing isolates, the Vitek ESBL test was compared with the NCCLS ESBL test and the DDST. The ESBL producer was defined as an organism showing an increase in the inhibited zone diameter of >or=5 mm for either cefotaxime or ceftazidime in combination with clavu-lanic acid versus its single test. The DDST was performed with 20 mm and 30 mm for interdisk diam-eter. For seven false negative isolates in the Vitek ESBL test, the DDST of cefepime was performed. RESULTS: Compared with the NCCLS ESBL test, the Vitek ESBL test showed one false positive (specificity, 99.6%), seven false negatives (sensitivity, 89.7%) and 97.5% agreement. Seven false negative isolates of the Vitek ESBL test were the cefoxitin-resistant ESBL producer. In positivity for the NCCLS ESBL test of 68 ESBL producing isolates, cefotaxime-clavulanic acid and ceftazidime-clavulanic acid were 94% and 91%. Cefotaxime, ceftazidime, aztreonam and ceftriaxone showed 95/90%, 100/55%, 100/85% and 95/80% positivity in double-disk synergy with amoxicillin-clavulanic acid (AMC) for 20/30 mm of the interdisk diameter respectively. For seven false negative isolates of the Vitek ESBL test, cefepime showed a distinct synergic effect with AMC. CONCLUSIONS: The Vitek ESBL test may be a useful method for clinical laboratories due to its easy, rapid and sensitive method but its method was less sensitive to cefoxitin-resistant ESBL. For these cases, if the NCCLS ESBL test or DDST with cefepime are added, the detection rate of the ESBL pro-ducer can be augmented.
Amoxicillin-Potassium Clavulanate Combination ; Aztreonam ; Cefotaxime ; Ceftazidime ; Ceftriaxone ; Diffusion ; Disease Outbreaks ; Escherichia coli* ; Escherichia* ; Klebsiella oxytoca ; Klebsiella*

BACKGROUND: The HBV-PCR assay seems to be potentially valuable diagnostic tool for the evaluation of variable serologic status. However, the selection of the primer for HBV-PCR test may be very important because they can influence the HBV-PCR positivity. METHODS: We compared the results of primer HBV1/2 including famous 1896 and 1899 mutation sites with those of primer PHBV1/2 at precore/core region. HBV-PCR was tested in 87 HBsAg-positive patients using two sets of primers. The results were evaluated according to the primers and also compared the results with the clinical diagnosis and the alanine aminotransferase (ALT) level. RESULTS: The positive rate of PHBV primer was higher than HBV primer including mutation sites (nucleotide 1896 and 1899) in HBeAg-negative patients. According to the clinical diagnosis, the sensitivity of PHBV primer was higher than that of HBV primer in chronic hepatitis patients. There was no significant correlation between ALT level and HBV-PCR results. CONCLUSIONS: It is important that the selection of primer in HBV-PCR is important, because the primer including mutation sites may result in false negative results. PHBV primer used in this study could be useful for the detection of HBV-DNA by HBV-PCR.
Alanine Transaminase ; Diagnosis ; Hepatitis, Chronic ; Humans ; Polymerase Chain Reaction*