Infection with Scopulariopsis brevicaulis after Cosmetic Surgery of the Face / 대한진단검사의학회지

Scopulariopsis brevicaulis is a ubiquitous soil saprophyte that commonly causes onychomycosis, accounting for 1-10% of such infections. Rarely, it may be responsible for cutaneous lesions or more severe infections, especially after traumatic or surgical injuries. We report of a 54-year-old female patient who developed facial cellulitis caused by S. brevicaulis, which occurred one year after the patient underwent cosmetic surgery of the face. The patient suffered from febrile sense, pain and a growing mass lesion on her left cheek, which were diagnosed as facial cellulitis associated with foreign material that had been implanted at the time of cosmetic surgery. Three pus cultures from the mass lesion which performed at a week interval yielded the same S. brevicaulis. Surgical removal and drainage by using liposuction procedure resulted in a favorable outcome. To our knowledge this is the first report of S. brevicaulis infection associated with cosmetic surgery in Korea.

A Simplified ABO Genotyping by Allele-Specific Polymerase Chain Reaction / 대한진단검사의학회지

BACKGROUND: Genotyping of ABO gene could be more informative and valuable than serological typing in some situations such as the resolution for ABO discrepancy between the cell typing and serum typing and determination of A and B subgroups. We developed a simple allele-specific polymerase chain reaction (AS-PCR) method without the use of any restriction enzymes to detect the A, B, O, and cis-AB alleles for Koreans. METHODS: An AS-PCR was designed with amplification refractory mutation system (ARMS) at nt (nucleotide) 261 (exon 6) and at nt 526, 803 (exon 7) of ABO gene to detect specific nucleotide sequence differences between the ABO alleles. We tested for ABO genotyping 60 DNA samples previously tested by PCR-RFLP and stored at -70degreeC. These samples had been obtained from blood donors recruited at the Gwangju-Chonnam Red Cross Blood Center between July 2002 and February 2003. RESULTS: With our new PCR method, the genotypes of the 60 samples were found to be A/O (n=10), A/A (n=5), B/O (n=10), B/B (n=5), O/O (n=10), cis-AB/A (n=5), cis-AB/B (n=5), and cis-AB/ O (n=10), which were the s ame results obtained previously with PCR-RFLP. CONCLUSIONS: Our AS-PCR is a simple and accurate method for the detection of A, B, O, and cis-AB alleles for Koreans.

A Case of Disseminated Trichosporon beigelii Infection Treated with the Combination of Amphotericin B and Fluconazole / 대한임상미생물학회지

Trichosporon beigelii is often resistant to the fungicidal effect of amphotericin B and can cause fatal disseminated infections in immunocompromised patients. We report a case of a disseminated T. beigelii infection with a favorable outcome in a patient with acute erythroleukemia and neutropenia. The patient presented a persistent fever, multiple erythematous skin lesions, and pulmonary infiltrates. T. beigelii was isolated from blood cultures in four days and also from cultures of abdominal skin lesion, sputum, and stool. The isolate was resistant to amphotericin B (MIC, 2 microgram/mL), and the respective fluconazole and itraconazole MICs were 4 and 1 microgram/mL. The patient was successfully treated with fluconazole plus amphotericin B in combination with granulocyte colony stimulating factor and leukocyte transfusion. This case shows the importance of early diagnosis and treatment with combination of amphotericin B and fluconazole as a prognostic factor of disseminated T. beigelii infections.

A Case of Bloodstream Infection Due to Fusarium oxysporum / 대한임상미생물학회지

Fusarium species are representative of the emerging group of filamentous molds, which cause respiratory and disseminated infections in immunocompromised patients. To date, only five cases of respiratory or disseminated skin infections due to Fusarium spp. have been described in Korea. Here we describe a fungemia case of Fusarium oxysporum in a 3-year old boy who was neutropenics following chemotheray for leukemia. Fever, painful macules on both extremities and phlebitis on the site of venous blood sampling developed on the day 35 of admission. All four blood cultures obtained on hospital days 37, 38, 40 and 42 yielded the same F. oxysporum. The infection was cured with a high dose (1.5 mg/kg) of amphotericin B. This case shows that Fusarium is among a few filamentous fungi that cause clinically detectable fungemias in immuncompromised hosts.

Clinical and Laboratory Features of Candidemia Caused by Different Candida Species / 대한진단검사의학회지

BACKGROUND: Recently, the incidence of candidemia due to Candida species other than C. albicans have increased. In this study, we analyzed the laboratory and clinical characteristics of candidemia caused by four different Candida species (C. albicans, C. parapsilosis, C. tropicalis and C. glabrata) occurring at Chonnam National University Hospital (CNUH). METHODS: The demographic, clinical and microbiological data of 157 patients with candidemia at CNUH from 1996 to 2002 was analyzed, retrospectively. The etiologic agents for 157 cases of candidemia were C. albicans (n=48), C. parapsilosis (n=48), C. tropicalis (n=32) and C. glabrata (n= 29). The characteristics of candidemia due to each single Candida species were compared with those with all other species combined. RESULTS: Although the majority (77%) of candidemic patients were adults, candidemia due to C. albicans or C. parapsilosis occurred significantly more often in premature infants (15%, retrospectively, P=0.002), in comparison with other Candida species (0%). Candidemia due to C. glabrata was more common in patients with neutropenia (41%, P<0.001), and they also occurred frequently in the absence of central venous catheter related candidemia (86%, P<0.001). Bloodstream infections with C. parapsilosis were more frequently the cause of catheter related candidemia (56%, P=0.012), and they had a better clinical outcome (90%, P=0.004) than those with other Candida species. CONCLUSIONS: This study confirms that some characteristics of candidemia such as age, underlying conditions, relatedness of catheter, and outcome can be different according to the species of Candida.

Correction Algorithm of Pseudothrombocytopenia due to Platelet Clumping / 대한진단검사의학회지

BACKGROUND: Pseudothrombocytopenia is a phenomenon that automated hematology analyzers calculate platelets at a spuriously low count. The most common cause of this phenomenon is platelet clumping. Several methods such as vortex mixing, changing anticoagulant to sodium citrate or heparin, and adding amikacin to ethylenediaminetetraacetic acid (EDTA) have been routinely used for managing pseudothrombocytopenia. The purposes of this study were to compare the efficacy of these four methods and to propose a cost-effective algorithm for managing pseudothrombocytopenia in clinical laboratories. METHODS: Ten patients (six males and four females) having pseudothrombocytopenia were evaluated. In these patients, platelet clumpings had been detected on more than three occasions by Coulter STKS (Beckman-Coulter, USA) and by microscopic examination on peripheral blood smear (PBS). We recollected blood samples from each patient in four tubes coated with EDTA, sodium citrate, heparin, or EDTA with amikacin. CBC of the blood samples in each tube was performed within one hour of collection; the samples in EDTA-coated tube were retested after vortex mixing. RESULTS: Platelet counts were increased in all cases (100%) by EDTA with amikacin as an anticoagulant, 80% (8/10) by vortex mixing or heparin, and in 90% (9/10) by sodium citrate. However, platelet counts were decreased in 20% (2/10) of heparin coated samples. `Clinically meaningful increase' was achieved in 60% (6/10) by heparin and EDTA with amikacin, in 50% (5/10) by sodium citrate, and in 40% (4/10) by vortex mixing. But `clinically meaningful decrease' was found in 10% (1/10) by heparin. CONCLUSIONS: When pseudothrombocytopenia due to platelet clumpings is detected, vortex mixing is recommended first. If platelet count does not increase after the vortex mixing, changing anticoagulant to sodium-citrate or adding amikacin to EDTA is recommended for managing pseudothrombocytopenia.

DNA Fingerprinting of Candida albicans Strains Isolated from Candidemic Patients by Polymerase Chain Reaction and Southern Hybridization Methods

BACKGROUND: Although several molecular typing methods have been used to investigate C. albicans infections, there remains no "gold standard" method by which relatedness of C. albicans strains is determined. In this study, two DNA fingerprinting methods were compared for genotyping of clinical strains of C. albicans isolated from candidemic patients. MATERIALS AND METHODS: Twenty-nine strains of C. albicans isolated from various clinical specimens (14 from blood, 7 from catheter, 4 from respiratory tract secretion, and 4 from urine) of 14 candidemic patients were analyzed. Primer 1245 and 1246 were employed for IR PCR and Southern blot hybridization method was used for C2 fingerprinting, with Ca3 and C1 as primers, after the fragmentation of DNA with EcoR1 RESULTS: IR PCR method separated 29 isolates into 9 (1245 primer), 7 (1246 primer) and 14 (combination of two primers) types, whereas C1 fingerprinting identified 16 different types. By combining the IR PCR and C1 fingerprinting methods, total of 16 different genotypes were identified among 29 isolates from 14 patients, which is the same result obtained by the C1 fingerprinting only. Using both methods, blood and non-blood isolates from each patient produced identical genotypes for 10 patients and different genotypes for 1 patient. In three patients, isolates from blood and other site of each patient showed identical patterns by IR PCR fingerprinting, but appeared different (n=1) or similar (n=2) by C1 fingerprinting. Overall, for 87% (13/15) of patients, isolates collected from catheter (6 of 7 patients), urine (4 of 4 patients), or respiratory (3 of 4 patients) were identical or similar to the corresponding blood isolates. CONCLUSION: Our study shows that C1 fingerprinting method is more discriminatory than IR PCR for the molecular typing of C. albicans isolates. For the majority of patients, blood and other site isolates had identical or similar genotypes.

DNA Fingerprinting of Candida albicans Strains Isolated from Candidemic Patients by Polymerase Chain Reaction and Southern Hybridization Methods

BACKGROUND: Although several molecular typing methods have been used to investigate C. albicans infections, there remains no "gold standard" method by which relatedness of C. albicans strains is determined. In this study, two DNA fingerprinting methods were compared for genotyping of clinical strains of C. albicans isolated from candidemic patients. MATERIALS AND METHODS: Twenty-nine strains of C. albicans isolated from various clinical specimens (14 from blood, 7 from catheter, 4 from respiratory tract secretion, and 4 from urine) of 14 candidemic patients were analyzed. Primer 1245 and 1246 were employed for IR PCR and Southern blot hybridization method was used for C2 fingerprinting, with Ca3 and C1 as primers, after the fragmentation of DNA with EcoR1 RESULTS: IR PCR method separated 29 isolates into 9 (1245 primer), 7 (1246 primer) and 14 (combination of two primers) types, whereas C1 fingerprinting identified 16 different types. By combining the IR PCR and C1 fingerprinting methods, total of 16 different genotypes were identified among 29 isolates from 14 patients, which is the same result obtained by the C1 fingerprinting only. Using both methods, blood and non-blood isolates from each patient produced identical genotypes for 10 patients and different genotypes for 1 patient. In three patients, isolates from blood and other site of each patient showed identical patterns by IR PCR fingerprinting, but appeared different (n=1) or similar (n=2) by C1 fingerprinting. Overall, for 87% (13/15) of patients, isolates collected from catheter (6 of 7 patients), urine (4 of 4 patients), or respiratory (3 of 4 patients) were identical or similar to the corresponding blood isolates. CONCLUSION: Our study shows that C1 fingerprinting method is more discriminatory than IR PCR for the molecular typing of C. albicans isolates. For the majority of patients, blood and other site isolates had identical or similar genotypes.

Plating Efficiency of Single CD34+ Cell from Bone Marrow, Peripheral Blood and Cord Blood / 대한혈액학회지

BACKGROUND: Although bone marrow (BM) CD34+cells and peripheral blood (PB) CD34+ cells are developmentally and functionally related, the recent data suggested that they may have different functional and clinical capabilities. Moreover, they have differential gene expression underlying the functional distinctions of primary human CD34+ hematopoietic stem and progenitor cells from BM and PB. The aim of this study was to investigate the plating efficiency of single CD34+ progenitor cell from BM, PB and umbilical cord blood (UCB). METHODS: After sorting, single CD34+ cells were cultured in individual wells of 96-well plates in serum-free medium containing selected hematopoietic growth factors, with or without G-CSF. Plating efficiency was microscopically determined by the presence of clusters of viable cells:[the number of positive (cells were present) wells/total wells]x100. CD34+ cell-derived colonies were classified according to the cell number per well. RESULTS: Although there was some variation of plating efficiency of CD34+ cells among six normal BMs, six PBs and five UCBs, overall average plating efficiency of single CD34+ cells from BM, PB and UCB was 30% (30.0+/-11.7, mean+/-SD), 79% (78.6+/-11.7) and 45% (45.3+/-9.3) respectively. As expected, the colony size was increased in the presence of G-CSF. CONCLUSION: The results of this study clearly showed that the different ex vivo expansion of single CD34+ progenitor cells from BM, PB and UCB. These might be an important data for understanding stem cell expansion in vivo and designing clinical application.

Correction of Platelet Count Using a Vortex in Pseudothrombocytopenia / 대한진단검사의학회지

BACKGROUND: Platelet clumping is a common cause of erroneous platelet counts by automated blood cell counter. The most commonly employed solution to this problem is to redraw the specimen into a different anticoagulant. However, this is unpleasant for the patient and not rapid for reporting of the corrected platelet count. Mixing of blood with a vortex mixer was evaluated as a method to disaggregate platelet clumps in blood and thus obtain accurate platelet counts. METHODS: Whole blood samples coated with ethylenediaminetetraacetic acid (EDTA) from 28 patients with platelet clumping and 20 controls without platelet clumping from July to September 2002 were mixed for 30 seconds with a vortex mixer. Platelet counts, blood smears, erythrocyte counts, Hgb, MCV and total leukocyte counts were evaluated before and after mixing. RESULTS: Vortex mixing of blood samples with platelet clumps caused an increased platelet count in 96% (27/28) and a decreased total leukocyte count in 68% (19/28). The mean platelet and total leukocyte counts of 28 blood samples before mixing were 155.0+/-89.6 (x10(3)/microL) and 12.9+/-5.5 (x10(3)/microL) and after mixing they were 249.2+/-116.2 (x10(3)/microL) and 12.0+/-5.4 (x10(3)/microL). Total erythrocyte counts, Hgb, MCV were not significantly affected by vortex mixing. Further, vortex mixing of 20 control samples had no consistent effect on each items. CONCLUSIONS: Vortex mixing of blood samples is a simple, rapid method without re-sampling in correction of erroneous platelet count induced by platelet clumps.

Two cases of partial-D showing different reactivity to various anti-D reagents / 대한수혈학회지

The 19-year-old twin sisters donated their blood in 2000. Their blood had typical Rh D negative red cell phenotype in Rh typing and weak D test using an anti-D reagent (Dade Behring, USA). Twin sisters donated blood again in 2001. Both were negative in anti-D reagent (Bioscotte Ltd., UK) and weakly positive in additively performed weak-D test. So we have acquired blood samples from them for further study in 2002. The red blood cells from twin sisters were not agglutinated with 4 various commercially available anti-D reagents. But in subsequently performed weak-D test, different reactivity to their anti-D reagents were shown, namely negative (Dade Behring, USA) and weakly positive (Ortho-clinical diagnostics, USA; Greencross, Korea; Bioscotte Ltd., UK). The lack of reactivity with some anti-D as shown in these cases can indicate the presence of a partial D antigen. So we carried out a additional serologic test using 6 monoclonal anti-D antibodies in partial-D typing set (Diamed, Switzerland) on Rh D antigens of red cells from twin sisters. According to the different reactivity patterns, we confirmed elder sister was partial-D category DFR and younger sister was partial-D with indeterminate category.