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Evaluation of Abbott Fourth Generation HIV Antigen and Antibody Assays.

Hee Jung KANG ; Kyeong Ha YOO ; Han Sung KIM ; Hyoun Chan CHO

The Korean Journal of Laboratory Medicine.2006;26(1):39-44. doi:10.3343/kjlm.2006.26.1.39

BACKGROUND: In order to reduce the diagnostic window period between the time of human immunodeficiency virus (HIV) infection and serological diagnosis, new fourth generation screening assays which detect HIV p24 antigen and specific antibody simultaneously have been developed. In this study, we evaluated the performance of a new fourth generation assay. METHODS: We compared a new fourth generation assay, Architect HIV Ag/Ab combo, with another fourth generation assay AxSYM HIV Ag/Ab combo and a third generation assay, AxSYM HIV 1/2 gO for their performance. The assays were evaluated using 3 HIV seroconversion panels, 305 sera of healthy subjects and 100 sera of patients with HBsAg or anti-HCV antibodies. Within-run and total coefficient variations of the three screening assays were analyzed for the evaluation of precision. RESULTS: Architect HIV Ag/Ab combo shortened the window period by 8.7+/-2.1 days relative to AxSYM HIV 1/2 gO and 2.0+/-2.0 days relative to AxSYM HIV Ag/Ab combo in seroconversion panels. Architect HIV Ag/Ab combo presented the best performance in precision among the three reagents; total CV for positive control was 3.6%, 9.6% and 4.6% for Architect HIV Ag/Ab combo, AxSYM HIV Ag/Ab combo and AxSYM HIV 1/2 gO, respectively. Specificities of three assays were not different in this study. CONCLUSIONS: HIV Ag/Ab combined assays reduced the diagnostic window as compared to the third generation screening assays, enabling an earlier diagnosis of HIV infection. A new fourth generation assay, Architect HIV Ag/Ab combo presents a better performance than AxSYM HIV Ag/Ab combo, showing improved seroconversion sensitivity and precision.
Diagnosis ; Hepatitis B Surface Antigens ; Hepatitis C Antibodies ; HIV Core Protein p24 ; HIV Infections ; HIV Seropositivity ; HIV* ; Humans ; Indicators and Reagents ; Mass Screening

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Endogenous Endophthalmitis by Aspergillus in a Patient with Multiple Myeloma.

Youn Hee PARK ; Chang Ki KIM ; Hyukmin LEE ; Kyong Ho ROH ; Jun Won JUNG ; Dongeun YONG ; Kyungwon LEE

The Korean Journal of Laboratory Medicine.2006;26(1):36-38. doi:10.3343/kjlm.2006.26.1.36

A 40-year-old man who had been treated for multiple myeloma, complained of decreased visual acuity of the left eye on the 30th day of admission. The nucleotide sequences of a fungal PCR product from vitreous fluid showed 99% homology with Aspergillus fumigatus (AY373851). Aspergillus spp. was isolated from vitreous fluid culture, also. Rapid diagnosis and intervention are critical elements for the Aspergillus endophthalmitis; therefore, it would be helpful to combine the fungal PCR with conventional fungus culture for clinically indicated specimens.
Adult ; Aspergillus fumigatus ; Aspergillus* ; Base Sequence ; Diagnosis ; Endophthalmitis* ; Fungi ; Humans ; Multiple Myeloma* ; Polymerase Chain Reaction ; Visual Acuity

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Infection with Scopulariopsis brevicaulis after Cosmetic Surgery of the Face.

Bong Joon OH ; Myong Jong CHAE ; Duck CHO ; Seung Jung KEE ; Myung Geun SHIN ; Jong Hee SHIN ; Soon Pal SUH ; Dong Wook RYANG

The Korean Journal of Laboratory Medicine.2006;26(1):32-35. doi:10.3343/kjlm.2006.26.1.32

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.
Cellulitis ; Cheek ; Drainage ; Female ; Humans ; Intraoperative Complications ; Korea ; Lipectomy ; Middle Aged ; Onychomycosis ; Scopulariopsis* ; Soil ; Suppuration ; Surgery, Plastic*

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Characterization of a Toxin A-Negative, Toxin B-Positive Variant Strain of Clostridium difficile.

Bo Moon SHIN ; Eun Young KUAK

The Korean Journal of Laboratory Medicine.2006;26(1):27-31. doi:10.3343/kjlm.2006.26.1.27

BACKGROUND: Clostridium difficile is one of the most important pathogens responsible for nosocomial diarrhea. Recently, we have frequently experienced culture positive, toxin A enzyme immunoassay negative strains. Therefore, we evaluated the strains with several PCR primer sets to characterize them. METHODS: A total of 351 stool specimens were examined for toxin A using enzyme linked fluorescent immunoassay (ELFA) and also cultured for C. difficile using cycloserine cefoxitine fructose agar incubated under anaerobic conditions. Spore stain and Vitek ANA identification card (BioMerieux, France) were used for identification of C. difficile. We amplified toxin A and toxin B genes in 81 isolates using primers NK1- NK2, NK3-NK2, NK9- NK11, and NK104-NK105. RESULTS: The concordance rate between ELFA and culture was 65.2% (229/351). PCR for the toxin A gene using NK1-NK2, NK3-NK2 and for the toxin B gene using NK104-NK105 showed almost the same results. However, toxin A gene PCR using NK9-NK11 showed that 45.7% (37/81) of the evaluated strains were toxin A (-)/ toxin B(+) variant strains; thus, the corrected sensitivity and specificity of the ELFA based on the PCR results for toxin A and B genes were 65.6% and 100%, respectively. CONCLUSIONS: The low sensitivity of the ELFA results for toxin A was due to the toxin A(-)/toxin B(+) variants of C. difficile, suggesting that the prevalence of the variant strains could be higher in Korea than was expected.
Agar ; Cefoxitin ; Clostridium difficile* ; Clostridium* ; Cycloserine ; Diarrhea ; Fructose ; Genes, vif ; Immunoassay ; Immunoenzyme Techniques ; Korea ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity ; Spores

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Emergence of CTX-M-12 and A Novel CTX-M Type Extended-Spectrum beta-Lactamaseproducing Klebsiella pneumoniae.

Il Kwon BAE ; Seok Hoon JEONG ; Kyungwon LEE ; Dongeun YONG ; Jongwook LEE ; Seong Geun HONG ; Eui Chong KIM ; Yeon Jun PARK ; Jung Oak KANG ; Young UH ; Jong Hee SHIN ; Wee Gyo LEE ; Ji young AHN ; Sung Hee LEE ; Gun Jo WOO ; Hyo Sun KWAK

The Korean Journal of Laboratory Medicine.2006;26(1):21-26. doi:10.3343/kjlm.2006.26.1.21

BACKGROUND: The aims of this study were to survey the nation-wide susceptibilities of Klebsiella pneumoniae isolates against ceftazidime and cefotaxime and to determine the prevalence of class A extended-spectrum beta-lactamases (ESBLs). METHODS: During the period of February to July 2004, K. pneumoniae isolates intermediate or resistant to ceftazidime and/or cefotaxime were collected from 12 hospitals in Korea. Antimicrobial susceptibilities were determined by the disk diffusion and the agar dilution methods and ESBL-production was by double-disk synergy test. Ceftazidime or cefotaxime-resistance determinants of the ESBLproducers were transfered to Escherichia coli J53 by transconjugation. Searches for class A ESBL genes were performed by PCR amplication. RESULTS: Among 212 clinical K. pneumoniae isolates, 172 (81%) isolates showed positive results in double-disk synergy test; the most prevalent ESBL was SHV-12 (n=104). Genes encoding ESBLs including SHV-2 (n=6), SHV-2a (n=17), CTX-M-3 (n=18), CTX-M-9 (n=6), CTX-M-12 (n=1), CTX-M- 14 (n=27), CTX-M-15 (n=3), and a novel CTX-M-type beta-lactamases were also detected. CONCLUSIONS: It is concluded that diversity of ESBLs in K. pneumoniae isolates are increasing in Korea. CTX-M-12 has never been reported in Asia, and a novel CTX-M-type ESBL has emerged.
Agar ; Asia ; beta-Lactamases ; Cefotaxime ; Ceftazidime ; Diffusion ; Escherichia coli ; Klebsiella pneumoniae* ; Klebsiella* ; Korea ; Pneumonia ; Polymerase Chain Reaction ; Prevalence

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Prevalence of Class A Extended-Spectrum beta-Lactamases in Clinical Isolates of Acinetobacter baumannii and Pseudomonas aeruginosa.

Se Jin OH ; Sang Uk LEE ; Hyun Yong HWANG ; Il Kwon BAE ; Hyun Soo JO ; Byung Ho LEE ; Seok Hoon JEONG

The Korean Journal of Laboratory Medicine.2006;26(1):14-20. doi:10.3343/kjlm.2006.26.1.14

BACKGROUND: Prevalence of class A extended-spectrum beta-lactamases (ESBLs) has been investigated repeatedly in members of family Enterobacteriaceae in Korea, but only rarely in Acinetobacter baumannii and Pseudomonas aeruginosa. The aims of this study were to determine the prevalence of class A ESBL-producing A. baumannii and P. aeruginosa and to characterize the genotypes. METHODS: During the period of June to September 2004, clinical isolates of A. baumannii and P. aeruginosa were collected from patients in Kosin University Gospel Hospital, Busan, Korea. Antimicrobial susceptibility was determined by the disk diffusion and the agar dilution methods, and ESBLproduction by the double-disk synergy test. Transferability of ceftazidime-resistance of ESBL-producers were tested by conjugation. The isoelectric points of ESBLs were determined by isoelectric focusing. Searches for blaTEM, blaSHV, blaCTX-M, blaPER-1, blaVEB, and blaGES/IBC genes were performed by PCR amplification, and the genotypes of ESBLs were determined by a direct nucleotide sequence analysis of the amplified products. RESULTS: A total of 58 clinical isolates of A. baumannii and 77 P. aeruginosa were collected. Three (5.2%) isolates of A. baumannii and four (5.2%) P. aeruginosa isolates showed positive results in the double-disk synergy test using ceftazidime and imipenem disks, and one (1.7%) A. baumannii and two (2.6%) P. aeruginosa isolates showed positive results in that test using ceftazidime and cefoxitin disks. The most prevalent class A ESBL genotype in A. baumannii isolates was blaPER-1 (n=6), and blaSHV-12 gene was also found in one P. aeruginosa isolate. CONCLUSIONS: It is concluded that class A PER-1 ESBL-producing A. baumannii isolates are spreading, and SHV-12-producing P. aeruginosa has emerged in Korea. The spread of class A ESBLs could compromise the future usefulness of expanded-spectrum -lactam antibiotics for the treatment of A. baumannii and P. aeruginosa infections.
Acinetobacter baumannii* ; Acinetobacter* ; Agar ; Anti-Bacterial Agents ; Base Sequence ; beta-Lactamases* ; Busan ; Cefoxitin ; Ceftazidime ; Diffusion ; Enterobacteriaceae ; Genotype ; Humans ; Imipenem ; Isoelectric Focusing ; Isoelectric Point ; Korea ; Polymerase Chain Reaction ; Prevalence* ; Pseudomonas aeruginosa* ; Pseudomonas*

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Detection of Enterovirus in Cerebrospinal Fluid by Real-Time Nested Reverse Transcription Polymerase Chain Reaction.

Se Ran HEO ; Sun Kyung JIN ; Ho Eun CHANG ; Kyoung Un PARK ; Junghan SONG ; Eui Chong KIM

The Korean Journal of Laboratory Medicine.2006;26(1):9-13. doi:10.3343/kjlm.2006.26.1.9

BACKGROUND: Enterovirus is a common cause of aseptic meningitis, respiratory disease and nonspecific febrile illness. The conventional methods for laboratory diagnosis of enterovirus infections have been virus culture and serotyping by an immunofluorecent test. We studied a new and more rapid approach for enterovirus detection in cerebrospinal fluid (CSF) by real-time nested PCR. METHODS: This study was performed on 50 CSF specimens from patients suspected of aseptic meningitis. Enterovirus was detected in CSF by PCRs for 3 different targets and real-time nested PCR. Enterovirus culture was also performed in 44 CSF specimens. RESULTS: The positive rate of PCRs for each of the 3 different targets was 26.0%, 40.0%, or 46.0%, and that of real-time nested PCR was 86.0%. Only 6.8% were positive in culture. Thus, the positive rate of real-time nested PCR was much higher than other methods. CONCLUSIONS: Our study revealed that the real-time nested PCR should be useful for diagnosis of enterovirus infections because of a high sensitivity and rapid detection.
Cerebrospinal Fluid* ; Clinical Laboratory Techniques ; Diagnosis ; Enterovirus Infections ; Enterovirus* ; Humans ; Meningitis, Aseptic ; Polymerase Chain Reaction* ; Real-Time Polymerase Chain Reaction ; Reverse Transcription* ; Serotyping

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Evaluation of COSMOsensor Glucose Monitoring System.

Eun Hyung YOO ; Hyun Jung CHO ; Chang Seok KI ; Soo Youn LEE

The Korean Journal of Laboratory Medicine.2006;26(1):1-8. doi:10.3343/kjlm.2006.26.1.1

BACKGROUND: Glucometer is a most widely-used point-of-care testing (POCT) analyzer and plays an important role in diabetes management. We evaluated the performance of the recently developed glucometer, COSMOsensor (Cosmogenome Inc., Seoul, Korea), comparing it with three foreign-made glucometers. METHODS: COSMOsensor was evaluated for linearity, precision, comparison of method and analysing time as well as the effect of operator. Other glucometers, Accu-Chek inform (Roche Diagnostics LTD., Mannheim, Germany), Precision(TM)PCx (Abbott Laboratories, Bedford, MA, USA), and Sure- Step.Flexx (LifeScan Inc., Milpitas, CA, USA) were evaluated for the same categories according to NCCLS guidelines. RESULTS: All four glucometers showed a good linearity (r> or =0.9814) and the within-run and total-run coefficients of variation (CVs) were within 3.5%. A high correlation (r> or =0.9659) was also found between the glucometers and Hitachi 7600 (Hitachi Co., Tokyo, Japan) in the central laboratory. Although differences with the reference method were within an allowable range, all glucometers showed variable bias compared with the reference method. CONCLUSIONS: The COSMOsensor showed a good analytical performance in linearity, precision, and correlation with the reference method, when compared with other foreign-made glucometers. Its rapid turnaround time and easy operation are appropriate for diabetes management and a rapid POCT analyzer. All glucometers showed variable biases, which might be due to different calibration status.
Bias (Epidemiology) ; Blood Glucose ; Calibration ; Glucose* ; Seoul

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Defining an Optimal Number of Immunophenotypic Markers for Lineage Assignment of Acute Leukemias Based on the EGIL Scoring System.

Seung Tae LEE ; Hee Jin KIM ; Sun Hee KIM

The Korean Journal of Laboratory Medicine.2006;26(6):393-399. doi:10.3343/kjlm.2006.26.6.393

BACKGROUND: The lineage assignment in acute leukemias is critical in therapeutic decisions. Immunophenotyping by flow cytometry plays the main role in the lineage assignment; however, few studies have been done to determine the optimal set of markers. In this regard, we tried to find out the optimal first-line set of markers with a minimal compromise in its diagnostic sensitivity. MATERIALS AND METHODS: We retrospectively analyzed 321 cases of acute leukemias whose diagnoses were based on the EGIL (European Group for Immunological Classification of Acute Leukemia) scores. At our institution, flow cytometic analyses included 15 first-line markers and 4 additional second-line markers as needed, along with immunohistochemical stains. We performed simulational studies for the expected EGIL scores involving every possible combination of markers and analyzing the overall diagnostic sensitivities in each combination. RESULTS: The cytoplasmic antigens including MPO stain and CD79a stain contributed greatly to the lineage assignment. For a sensitivity over 95%, there needed a combination of MPO stain with other 5 flow markers (CD33, CD13, CD14, CD15 and CD117) for myeloid lineage; CD79a stain with 3 flow markers [CD19, CD10, and CD20 (or TdT)] for B-lymphoid lineage; and 4 flow markers (CD2, CD3, CD5, and CD7) for T-lymphoid lineage. CONCLUSIONS: To maintain diagnostic sensitivities over 95% for each lineage, at least 14 markers (including MPO stain and CD79a stain) were needed; while 16 markers were needed for a sensitivity of 100%. When combined with other important markers for specific aims such as CD45, the minimum number of markers needed for the accurate diagnosis of acute leukemias would be more than about 18 to 20.
Classification ; Coloring Agents ; Cytoplasm ; Diagnosis ; Flow Cytometry ; Immunophenotyping ; Leukemia* ; Retrospective Studies

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Inverse Tendency between Ex Vivo Expansion Potential of Hematopoietic Progenitors and Time to Engraftment after Hematopoietic Stem Cell Transplantation.

Ji Myung KIM ; Chan Jeoung PARK ; Hyun Sook CHI ; Jae Hwan LEE ; Gyu Hyung LEE ; Jong Jin SEO

The Korean Journal of Laboratory Medicine.2006;26(6):385-392. doi:10.3343/kjlm.2006.26.6.385

BACKGROUND: The CD34+ cell dose and infused number of committed progenitor cells in transplantation are important factors in hematologic engraftment. However, the relationship between expansion potential of progenitor cells and hematologic engraftment remains controversial. We evaluated whether expansion potential of progenitor cells is a predictive factor of post-transplantation hematologic engraftment. METHODS: Mononuclear cells isolated from mobilized peripheral blood and bone marrow were cultured with cytokine cocktail for 7 days. Progenitor cells and committed progenitors were analyzed using stem cell markers (CD34 and CD133) and lineage specific markers. Hematologic engraftment was defined as neutrophil counts over 500/microliter and platelet counts over 20,000/microliter without transfusion. Acute and chronic graft-versus-host disease (GVHD) were investigated. RESULTS: There was inverse tendency between the number and fold expansion of progenitor cells or committed (granulocytic or megakaryocytic) progenitors and time to engraftment. Especially, fold expansion of CD34(+)/CD33(+) cells was significantly correlated with time to neutrophil engraftment in bone marrow transplantation (r=-0.56, P=0.04). The infused number and fold expansion of lymphoid progenitors were not related to the occurrence of acute or chronic GVHD. CONCLUSIONS: We could not prove that expansion potential of progenitor cells and committed progenitor cells is correlated to hematologic engraftment although there is a correlation between CD34(+)/ CD33(+) cells and time to neutrophil engraftment. But, a further study on the value of expansion potential is required because there is an inverse tendency.
Bone Marrow ; Bone Marrow Transplantation ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Neutrophils ; Platelet Count ; Stem Cells

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