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

BACKGROUND: This study was carried out in order to find an efficient and reliable method for genotyping Serratia marcescens, which frequently causes a nosocomial infection, by PCR-based methods. METHODS: The DNA fingerprinting by RAPD, ERIC and REP-PCR was performed for 24 isolates of S. marcescens obtained from 24 patients hospitalized at Chungbuk National University Hospital from March to May 2001. Reproducibility of the methods and minimum inhibitory concentration (MIC) of antibiotics against the isolates were also determined. RESULTS: The patients bearing epidemic S. marcescens were catheterized in their urinary tracts and admitted to the ICU or neurosurgical ward. All of the epidemic isolates showed the same susceptibility pattern. The epidemic isolates demonstrated resistance to all the test antibiotics except imipenem. The non-epidemic isolates showed variable susceptibility with a low MIC, except for one isolate. The DNA fingerprints by RAPD, ERIC and REP-PCR demonstrated an identical band pattern in epidemic strains but demonstrated different band patterns in non-epidemic strains. The PCR-based methods were found to be easy to manipulate, rapid in reaching conclusions, powerful in discrimination, and reproducible in results. CONCLUSIONS: RAPD, ERIC-PCR and REP-PCR are proved to be a reliable and efficient method for genotyping the epidemic S. marcescens in the laboratory.
Anti-Bacterial Agents ; Catheters ; Chungcheongbuk-do ; Cross Infection ; Discrimination (Psychology) ; DNA Fingerprinting ; Epidemics ; Humans ; Imipenem ; Microbial Sensitivity Tests ; Molecular Typing* ; Serratia marcescens* ; Urinary Tract

BACKGROUND: VIM-2 type metallo-beta-lactamase (MBL) producing strains are presently spreading to Pseudomonas spp., Acinetobacter spp. and even to Enterobacteriaceae such as Serratia marcescens, Enterobacter cloacae and Klebsiella pneumoniae in Korea. Recently we determined the phenotype and the genotype of three MBL-producing Providencia rettgeri isolated from urinary specimen of three patients with neurosurgical ward, and analyzed the blaVIM-2 containing integron of a P. rettgeri CBU852. METHODS: EDTA-disk synergy test was used for the screening of MBL, and the PCR for blaIMP-1, blaVIM-1 and blaVIM-2 was performed. The minimal inhibitory concentration of those isolates was determined by broth microdilution method, and the genomic DNA fingerprinting analysis was performed by random amplified polymorphic DNA (RAPD). The sequence of the blaVIM-2 containing integron was determined. RESULTS: Three P. rettgeri with reduced imipenem susceptibility showed the positive EDTA-disk synergy test and blaVIM-2 was detected by PCR. Antimicrobial susceptibility test showed the resistance to all beta-lactams tested, ciprofloxacin and aminoglycoside such as gentamicin, tobramycin and amikacin, indicating multidrug resistance of those isolates. RAPD analysis showed the identical DNA fingerprint of those three isolates. The novel class 1 integron, including aacA4, blaVIM-2, orf "ii" and orf "iii", was detected in a P. rettgeri CBU852. CONCLUSIONS: In this study, the multidrug resistant P. rettgeri CBU852 had blaVIM-2 containing novel class 1 integron. The emergence of blaVIM-2 producing P. rettgeri could compromise the use of carbapenem in treatment of infections caused by MBL producing bacteria. To our knowledge, this is the first report that VIM-2 MBL gene has been detected in P. rettgeri.
Acinetobacter ; Amikacin ; Animals ; Bacteria ; beta-Lactams ; Ciprofloxacin ; DNA ; DNA Fingerprinting ; Drug Resistance, Multiple ; Ecthyma, Contagious ; Enterobacter cloacae ; Enterobacteriaceae ; Genotype ; Gentamicins ; Humans ; Imipenem ; Integrons ; Klebsiella pneumoniae ; Korea ; Mass Screening ; Phenotype ; Polymerase Chain Reaction ; Providencia* ; Pseudomonas ; Serratia marcescens ; Tobramycin

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

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: Basic Fibroblast Growth Factor (bFGF) is known to be closely related to myelofibrosis and hematopoiesis including magakaryopoiesis. The main bone marrow finding in patients with idiopathic thrombocytopenic purpura (ITP) is an increased megakaryopoiesis without myelofibrosis. Purposes of this study are to evaluate the changes in bFGF expression pattern in the bone marrow of patients with ITP and to correlate them with the plasma concentrations of bFGF. METHODS: Paraffin-sections of bone marrow biopsies from 17 cases ITP and 7 cases normal controls, without pathological alterations, were investigated by immunohistochemistry for bFGF and CD68. The plasma levels of bFGF were evaluated by enzyme immunoassay in 7 cases of ITP and controls. RESULTS: The bFGF was strongly expressed in stromal cells and weakly in megakaryocytes in normal controls. The density of the bFGF-expressing stromal cells was decreased in 70% (12/17) of the patients with ITP, compared with none in the other controls. The number of stromal cells in patients with ITP was similar to those in the control groups. The bFGF plasma levels were significantly lower in almost all the ITP patients compared to the control group. CONCLUSIONS: The results indicate that concentrations of bFGF in plasma and bone marrow stromal cells of ITP were decreased. Although the mechanism of low cellular and plasma concentrations of bFGF needs to be elucidated, these findings may complement the serologic and morphological diagnosis of ITP.
Biopsy ; Bone Marrow* ; Complement System Proteins ; Diagnosis ; Fibroblast Growth Factor 2* ; Hematopoiesis ; Humans ; Immunoenzyme Techniques ; Immunohistochemistry ; Megakaryocytes ; Mesenchymal Stromal Cells ; Plasma* ; Primary Myelofibrosis ; Purpura, Thrombocytopenic, Idiopathic* ; Stromal Cells

PURPOSE: Nasal cytology for eosinophils has been reported to be very useful for the diagnosis of allergic rhinitis. The purpose of this study is to describe the relationship of the appearance of nasal eosinophils with the levels of total eosinophil counts, total IgE, and house dust mite specific IgE in child patients with symptoms of rhinitis. METHODS: Two hundred seventy eight children with symptoms of rhinitis less than 16 years of age were recruited and evaluated for the following variables: total eosinophil counts, total IgE concentrations, house dust mite specific IgE concentrations, and nasal cytology for eosinophils. RESULTS: The rate of appearance of nasal eosinophils graded as positive rose as the children's age increased. The levels of total eosinophil counts, total IgE concentrations and house dust mite-specific IgE concentrations were significantly higher in children with nasal eosinophils graded as positive than those with less than 5 percent of nasal eosinophils. The rates of appearance of nasal eosinophils graded as positive below and above 250/microL of total eosinophil counts, 250 kUa/L of total IgE concentrations, and 2 kUa/L of house dust mite (D. pteronyssinus or D. farinae) specific IgE concentrations were 16 and 41 percent, 27 and 56 percent, and 13 and 68 percent, respectively CONCLUSION: The levels of total eosinophil counts, total IgE concentrations, and house dust mite specific IgE concentrations correlate significantly with the recovery of nasal eosinophils in children with symptoms of rhinitis.
Child* ; Diagnosis ; Dust* ; Eosinophils* ; Humans ; Immunoglobulin E* ; Pyroglyphidae* ; Rhinitis*

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*

Toxic shock syndrome is an acute and febrile illness that rapidly progress to shock and multi-organ failure, and it is caused by toxin-producing strains of Staphylococcus aureus or Streptococcus species. Streptococcal toxic shock syndrome (STSS) is usually caused by group A streptococci, but non-group A STSS is rare. In this study, we describe a case of STSS caused by Streptococcus agalactiae(group B streptococci) in a patient with alcoholic liver cirrhosis. At arrival in our hospital, the patient had a decreased mental status with hemorrhagic bullae on four extremities, and he progressed to a fatal outcome within 4 days in spite of antibiotic treatment.
Extremities ; Fatal Outcome ; Humans ; Liver Cirrhosis ; Liver Cirrhosis, Alcoholic ; Shock ; Shock, Septic ; Staphylococcus aureus ; Streptococcus ; Streptococcus agalactiae