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1.
Ann Clin Lab Sci ; 46(5): 562-5, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27650627

RESUMO

A novel FLCN c.1489_1490delTG (p.Val497Glyfs*22) mutation at the genomic DNA and mRNA levels was identified in a 43-year-old woman with complaining of recurrent primary spontaneous pneumothorax. The aberrant FLCN mRNA escaped the nonsense-mediated decay system (NMD) because of a premature termination code located in an NMD-incompetent region. To the best of our knowledge, this is the first case report of an FLCN mutation escaping the NMD.


Assuntos
Mutação/genética , Degradação do RNAm Mediada por Códon sem Sentido/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Supressoras de Tumor/genética , Adulto , Sequência de Bases , Éxons/genética , Feminino , Humanos , Pneumotórax/diagnóstico por imagem , Pneumotórax/genética , Tomografia Computadorizada por Raios X
2.
Leuk Res ; 47: 172-7, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27379838

RESUMO

Micro (mi)RNA dysregulation is implicated in the development of myelodysplastic syndrome (MDS). Chromosomal abnormalities on 1q are frequently detected in Korean patients with MDS; however, how these are related to disease development is unknown. The present study compared the expression profiles of miRNAs encoded by chromosome 1q between 65 MDS patients and 11 controls. We found that miR-205-5p levels were 12.5 fold higher in the former (P=0.001). miR-205-5p level was increased in 44.7% of patients when an arbitrary 2(-ΔCt) cut-off value of 1.25 was used. miR-205-5p expression data were used to generate a receiver operating characteristic (ROC) curve for miR-205-5p, for which the area under the curve (AUC) was 0.825 (95% confidence interval: 0.710-0.941; P=0.001). Moreover, transfection with a miR-205-5p mimic induced cell proliferation by inhibiting the expression of the tumor suppressor protein phosphatase and tensin homolog (PTEN). Our findings suggest that miR-205-5p upregulation contributes to MDS by suppressing PTEN and that miR-205-5p thus acts as an oncogene in hematopoietic cells.


Assuntos
MicroRNAs/genética , Síndromes Mielodisplásicas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Linhagem Celular , Proliferação de Células , Aberrações Cromossômicas , Cromossomos Humanos Par 1/genética , Feminino , Perfilação da Expressão Gênica , Humanos , Cariotipagem , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/patologia , PTEN Fosfo-Hidrolase/antagonistas & inibidores , Curva ROC , República da Coreia , Regulação para Cima , Adulto Jovem
3.
Ann Clin Lab Sci ; 46(3): 270-8, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27312551

RESUMO

Molecular methods have the potential to improve the speed and accuracy of Acinetobacter species identification in clinical settings. The goal of this study is to develop species-specific PCR assays based on differences in the RNA polymerase beta-subunit gene (rpoB) to detect nine commonly isolated Acinetobacter species including Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter pittii, Acinetobacter nosocomialis, Acinetobacter lwoffii, Acinetobacter ursingii, Acinetobacter bereziniae, Acinetobacter haemolyticus, and Acinetobacter schindleri. The sensitivity and specificity of these nine assays were measured using genomic DNA templates from 55 reference strains and from 474 Acinetobacter clinical isolates. The sensitivity of A. baumannii-specific PCR assay was 98.9%, and the sensitivity of species-specific PCR assays for all other species was 100%. The specificities of A. lwoffii- and A. schindleri-specific PCR were 97.8 and 98.9%, respectively. The specificity of species-specific PCR for all other tested Acinetobacter species was 100%. The lower limit of detection for the nine species-specific PCR assays developed in this study was 20 or 200 pg of genomic DNA from type strains of each species. The Acinetobacter species-specific PCR assay would be useful to determine the correct species among suggested candidate Acinetobacter species when conventional methods including MALDI-TOF MS identify Acinetobacter only to the genus level. The species-specific assay can be used to screen large numbers of clinical and environmental samples obtained for epidemiologic study of Acinetobacter for the presence of target species.


Assuntos
Acinetobacter/genética , Acinetobacter/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Limite de Detecção , Sensibilidade e Especificidade , Especificidade da Espécie
4.
Ann Lab Med ; 36(2): 124-30, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26709259

RESUMO

BACKGROUND: Acinetobacter baumannii infections are difficult to treat owing to the emergence of various antibiotic resistant isolates. Because treatment options are limited for multidrug-resistant (MDR) A. baumannii infection, the discovery of new therapies, including combination therapy, is required. We evaluated the synergistic activity of colistin, doripenem, and tigecycline combinations against extensively drug-resistant (XDR) A. baumannii and MDR A. baumannii. METHODS: Time-kill assays were performed for 41 XDR and 28 MDR clinical isolates of A. baumannii by using colistin, doripenem, and tigecycline combinations. Concentrations representative of clinically achievable levels (colistin 2 µg/mL, doripenem 8 µg/mL) and achievable tissue levels (tigecycline 2 µg/mL) for each antibiotic were used in this study. RESULTS: The colistin-doripenem combination displayed the highest rate of synergy (53.6%) and bactericidal activity (75.4%) in 69 clinical isolates of A. baumannii. Among them, the-doripenem-tigecycline combination showed the lowest rate of synergy (14.5%) and bactericidal activity (24.6%). The doripenem-tigecycline combination showed a higher antagonistic interaction (5.8%) compared with the colistin-tigecycline (1.4%) combination. No antagonism was observed for the colistin-doripenem combination. CONCLUSIONS: The colistin-doripenem combination is supported in vitro by the high rate of synergy and bactericidal activity and lack of antagonistic reaction in XDR and MDR A. baumannii. It seems to be necessary to perform synergy tests to determine the appropriate combination therapy considering the antagonistic reaction found in several isolates against the doripenem-tigecycline and colistin-tigecycline combinations. These findings should be further examined in clinical studies.


Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Colistina/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Minociclina/análogos & derivados , Infecções por Acinetobacter/tratamento farmacológico , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/genética , Acinetobacter baumannii/isolamento & purificação , Antibacterianos/uso terapêutico , Proteínas de Bactérias/genética , Carbapenêmicos/uso terapêutico , Colistina/uso terapêutico , Doripenem , Sinergismo Farmacológico , Quimioterapia Combinada , Humanos , Testes de Sensibilidade Microbiana , Minociclina/farmacologia , Minociclina/uso terapêutico , Tipagem de Sequências Multilocus , Tigeciclina , beta-Lactamases/genética
5.
Microbiol Immunol ; 59(5): 285-98, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25786669

RESUMO

The four methods for assigning bacterial species are the Clinical and Laboratory Standards Institute (CLSI), modified CLSI (mCLSI), phylogenetic analysis (PA) and closest match (CM) methods, these are used to identify the genus and species using 16S rRNA gene sequence results. In this study, the results of identification by these four methods of 37 aerobic reference strains, 30 anaerobic reference strains, 15 Acinetobacter reference strains and 167 Acinetobacter clinical strains were compared. The rates of accurate identification to the species level using the CLSI, mCLSI, PA and CM methods were as follows: 24.3, 86.5, 86.5 and 89.2%, respectively, for the 37 aerobic reference strains; 73.3%, 96.7%, 90.0% and 93.3%, respectively, for the 30 anaerobic reference strains; 40.0%, 93.3%, 100% and 93.3%, respectively, for the 15 Acinetobacter reference strains; and 53.9%, 90.4%, 95.8% and 90.4%, respectively, for the 167 Acinetobacter clinical strains. The rates of accurate identification to the genus level using the CLSI, mCLSI, PA, and CM methods were as follows: 91.9%, 91.9%, 94.6% and 91.9%, respectively, for the 37 aerobic reference strains; 100%, 100%, 100% and 100%, respectively, for all of the 30 anaerobic reference strains, 15 Acinetobacter reference strains and the 167 Acinetobacter clinical strains. The mCLSI is the most practical and pragmatic method for identification of species based on 16S rRNA sequences for hospital, research or industry laboratories because it performs well and involves a simple procedure.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas/métodos , DNA Ribossômico/genética , Técnicas de Diagnóstico Molecular/métodos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Bactérias/classificação , Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , Genes de RNAr , Humanos
6.
Acta Haematol ; 133(1): 101-5, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25227135

RESUMO

8p11 myeloproliferative syndrome (EMS) is a rare disease characterized by myeloproliferative neoplasm (MPN) associated with eosinophilia and T or B lymphoblastic lymphoma/leukemia. EMS is defined by molecular disruption of the FGFR1 gene at the 8p11-12 chromosome locus, and various partner genes are associated with FGFR1 gene translocation or insertion. The different partner-FGFR1 fusion genes are associated with slightly different disease phenotypes. The present patient showed T lymphoblastic lymphoma in a cervical lymph node, involvement of malignant lymphoma in the skin, and MPN bone marrow morphology with peripheral monocytosis. Chromosome analysis of the patient showed t(1;8)(q25;p11.2). To our knowledge, only 2 cases of EMS with translocation of t(1;8)(q25;p11.2) have been previously reported. Including this case, all 3 cases with EMS with t(1;8)(q25;p11.2) showed MPN bone marrow morphology and peripheral monocytosis. These findings support that t(1;8)(q25;p11.2) is associated with peripheral monocytosis in EMS patients. Of the 2 cases of EMS with t(1;8)(q25;p11.2) which were previously reported, FGFR1 rearrangement was not confirmed in 1 case. Similarly, FGFR1 rearrangement in the present case was not detected by fluorescence in situ hybridization or reverse transcription-polymerase chain reaction. Further study is needed to identify other techniques that could be used to demonstrate FGFR1 rearrangement.


Assuntos
Cromossomos Humanos Par 1 , Cromossomos Humanos Par 8 , Transtornos Mieloproliferativos/genética , Translocação Genética , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Medula Óssea/patologia , Bandeamento Cromossômico , Humanos , Hibridização in Situ Fluorescente , Linfonodos/patologia , Masculino , Pessoa de Meia-Idade , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/tratamento farmacológico , Pele/patologia , Tomografia Computadorizada por Raios X , Resultado do Tratamento
7.
Pharmacogenomics ; 15(8): 1117-24, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25084204

RESUMO

BACKGROUND: NAT2 is a common metabolizer of many clinical drugs. NAT2 haplotyping requires a complex procedure. Allele-specific PCR followed by direct sequencing or cloning sequencing are common methods used for haplotyping. However, these common methods require labor-intensive procedures. Allele-specific sequencing was designed for haplotyping of the NAT2 gene. MATERIALS & METHODS: Using rapid DNA polymerase with high fidelity, we amplified the NAT2 coding region of genomic DNA for direct sequencing, allele-specific sequencing and for the cloning of genomic DNA from 307 healthy Korean subjects. Direct sequencing analysis of the 870-bp coding region of NAT2 was performed in order to search 11 of the most common SNPs. For cases who were heterozygous for two or more SNPs and whose haplotypes were not determined by direct sequencing, we performed sequencing analysis using the allele-specific sequencing primer for one specified allele. We performed cloning-sequencing analysis for confirmation of the haplotyping results of allele-specific sequencing. RESULTS: Homozygotes for SNPs, heterozygotes for one SNP and heterozygotes for two or more SNPs were 142 (46.3%), six (2.0%) and 259 (51.8%) cases, respectively. There was 100% concordance between the results of NAT2 haplotyping using allele-specific sequencing and cloning sequencing of 65 cases that were heterozygous for two or more SNPs in 307 samples. For cases that were homozygous for the SNPs by direct sequencing, the haplotypes of NAT2 were clearly determined by cloning sequencing. CONCLUSION: We have developed a novel method for NAT2 haplotyping using allele-specific sequencing, which could be an innovative and reliable method for NAT2 haplotyping.


Assuntos
Arilamina N-Acetiltransferase/genética , Haplótipos/genética , Polimorfismo de Nucleotídeo Único/genética , Análise de Sequência de DNA , Alelos , Povo Asiático/genética , Clonagem Molecular , Humanos
8.
Diagn Microbiol Infect Dis ; 78(1): 29-34, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24157058

RESUMO

Since accurate identification of species is necessary for proper treatment of Acinetobacter infections, we compared the performances of 4 bacterial identification methods using 167 Acinetobacter clinical isolates to identify the best identification method. To secure more non-baumannii Acinetobacter (NBA) strains as target strains, we first identified Acinetobacter baumannii in a total of 495 Acinetobacter clinical isolates identified using the VITEK 2 system. Because 371 of 495 strains were identified as A. baumannii using gyrB multiplex 1 PCR and blaOXA51-like PCR, we performed rpoB gene sequencing and 16S rRNA gene sequencing on remaining 124 strains belonging to NBA and 52 strains of A. baumannii. For identification of Acinetobacter at the species level, the accuracy rates of rpoB gene sequencing, 16S rRNA gene sequencing, gyrB multiplex PCR, and the VITEK 2 were 98.2%, 93.4%, 77.2%, and 35.9%, respectively. The gyrB multiplex PCR seems to be very useful for the detection of ACB complex because its concordance rates to the final identification of strains of ACB complex were 100%. Both the rpoB gene sequencing and the 16S rRNA gene sequencing may be useful in identifying Acinetobacter.


Assuntos
Infecções por Acinetobacter/diagnóstico , Acinetobacter/classificação , Acinetobacter/isolamento & purificação , Técnicas de Tipagem Bacteriana/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Acinetobacter/genética , Acinetobacter/metabolismo , Infecções por Acinetobacter/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , DNA Girase/genética , RNA Polimerases Dirigidas por DNA/genética , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Adulto Jovem
9.
Ann Clin Lab Sci ; 42(4): 370-4, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23090732

RESUMO

This study was designed to compare two automated systems and one manual system for hepatitis B virus (HBV) nucleic acid extraction. The two automated systems were the MagNA Pure 96 system (Roche Applied Science, Manheim, Germany) and the Chemagic system (Chemagen, Baesweiler, Germany), and the manual system was the QIAamp system (Qiagen, Hilden, Germany). Sixty-eight samples that were within the detection range of the Cobas Ampliprep/Cobas TaqMan (CAP/CTM) platform (Roche Molecular Systems, Manheim, Germany) were selected. Extracted viral nucleic acids from the three systems were quantified using an AccuPower HBV Quantitative PCR kit (Bioneer, Daejon, Korea). The MagNA Pure 96 system and QIAamp system did not detect viral loads in one sample. The Chemagic system did not detect low viral loads in nine samples (range, 26-290 IU/mL by the CAP/CTM platform). Comparisons of the viral loads of the samples from the MagNA Pure 96 system, the Chemagic system, and the QIAamp system with those from the CAP/CTM platform yielded correlation coefficients of 0.977, 0.914, and 0.967, respectively. Comparisons of the MagNA Pure 96 system and the Chemagic system with the QIAamp system yielded correlation coefficients of 0.987 and 0.939, respectively. The MagNA Pure 96 system demonstrated better performance than the Chemagic system for HBV nucleic acid extraction. The MagNA Pure 96 system demonstrated comparable performance with the QIAamp system.


Assuntos
Fracionamento Químico/métodos , Vírus da Hepatite B/química , Ácidos Nucleicos/isolamento & purificação , Carga Viral/instrumentação , Carga Viral/métodos , Reação em Cadeia da Polimerase em Tempo Real
10.
Jpn J Infect Dis ; 65(2): 162-6, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22446125

RESUMO

Between January 2004 and December 2004, an outbreak of imipenem-resistant Acinetobacter baumannii (IRAB) in 2 intensive care units (ICU) of Chosun University Hospital, Korea affected 77 patients. A case-control study revealed that the time spent in the hospital and mechanical ventilation practices were risk factors. IRAB was isolated from the hands of 4% (5/124) of healthcare workers; 27.3% (21/77) of the samples obtained from the ICU environment. A pulsed-field gel electrophoresis analysis showed that 82.1% (23/28) of clinical IRAB isolates and 85.7% (6/7) of environmental IRAB isolates were type A. The ISAba1F/OXA-51-likeR PCR showed that 93.7% (30/32) of IRAB strains had the ISAba1 gene upstream of the bla(OXA-51-like) gene. Two ISAba1F/OXA-51-likeR PCR-negative IRAB strains were bla(IMP-1) positive. All of the IRAB strains tested by PCR were negative for bla(VIM), bla(SIM), bla(GIM-1), bla(SPM-1), bla(GES), bla(OXA-23-like), bla(OXA-24-like), and bla(OXA-58-like) carbapenemase genes. After implementing an infection control strategy, a steady reduction in the attack rate of IRAB infection was observed.


Assuntos
Infecções por Acinetobacter/epidemiologia , Acinetobacter baumannii/classificação , Acinetobacter baumannii/efeitos dos fármacos , Surtos de Doenças , Imipenem/farmacologia , Resistência beta-Lactâmica , beta-Lactamases/genética , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/enzimologia , Acinetobacter baumannii/isolamento & purificação , Estudos de Casos e Controles , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Feminino , Genótipo , Mãos/microbiologia , Pessoal de Saúde , Hospitais , Humanos , Unidades de Terapia Intensiva , Coreia (Geográfico)/epidemiologia , Masculino , Epidemiologia Molecular , Tipagem Molecular , Fatores de Risco
11.
Ann Clin Lab Sci ; 41(4): 340-5, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22166503

RESUMO

ABO is the most clinically important blood group system in transfusion and transplantation medicine. The popular ABO genotyping methods, such as the sequencing of exons 6 and 7 and sequence-specific primer (SSP)-PCR, often lead to ambiguous typing results. Long PCR-sequencing method was designed to analyze two regulatory regions (promoter and CBF/NF-Y enhancer regions) and all genomic sequences (except for intron 1) of the ABO gene. Using rapid DNA polymerase with high-fidelity, we amplified 6.3 kb and 7.3 kb for sequencing of enhancer-exon 1 and exons 2-7, respectively. ABO genotyping was performed using this technique in the peripheral blood of three unrelated families. The time requirements of the PCR amplification and purification processes were about 2.0 hours and 15 minutes, respectively. Five different ABO alleles (ABO A102, ABO A105, ABO O01, ABO O02, and ABO B101) with allele-specific CBF/NF-Y minisatellite repeats from three families were analyzed. All genotyping results agreed with serologic findings and results expected by Mendelian inheritance. Compared to conventional PCR-direct sequencing for ABO genotyping, this method proves simple and fast for the analysis of ABO genotypes. Therefore, it might be valuable in clinical transfusion or forensic applications.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Técnicas de Genotipagem/métodos , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA/métodos , DNA/análise , DNA/genética , Feminino , Humanos , Masculino
12.
Diagn Microbiol Infect Dis ; 70(4): 442-7, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21767700

RESUMO

To compare the identification accuracies of VITEK 2 (bioMérieux), MicroScan (Siemens Healthcare), and Phoenix (Becton Dickinson), microbial identification was performed on 160 clinical isolates and 50 reference strains on each of these 3 systems, using the appropriate identification kit provided by each system. Of the 142 clinical isolates that were identified at the species level, VITEK 2, MicroScan, and Phoenix correctly identified 93.7%, 82.4%, and 93.0%, and incorrectly identified 2.1%, 7.0%, and 0%, respectively. In the reference strain tests, VITEK 2, MicroScan, and Phoenix correctly identified 55.3%, 54.4%, and 78.0% of the reference strains at the species level and incorrectly identified 10.6%, 13.0%, and 6.0% of the reference strains, respectively. In conclusion, the identification rate of VITEK 2, Phoenix, and MicroScan was high or acceptable on clinical isolates. Phoenix showed a significantly higher performance than VITEK 2 or MicroScan in identifying the reference strains.


Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas/métodos , Erros de Diagnóstico/estatística & dados numéricos , Humanos , Sensibilidade e Especificidade
13.
Korean J Lab Med ; 30(6): 567-74, 2010 Dec.
Artigo em Coreano | MEDLINE | ID: mdl-21157140

RESUMO

BACKGROUND: JAK2 genetic variations have been described in a high proportion of patients with BCR/ABL1-negative myeloproliferative neoplasms (MPN). This study was designed to analyze the frequencies of JAK2 V617F and exon 12 variations, and their correlations with clinical characteristics of Korean patients with BCR/ABL1-negative MPN. METHODS: We examined a total of 154 patients with BCR/ABL1-negative MPN that included 24, 26, 89, and 15 patients with polycythemia vera (PV), primary myelofibrosis (PMF), essential thrombocythemia (ET), and unclassified myeloproliferative neoplasms (MPNU), respectively. We performed allele-specific PCR to detect V617F in all BCR/ABL1-negative patients, and performed direct sequencing to detect exon 12 variations in 47 V617F-negative MPN patients. JAK2 c.1641+179_183del5 variation was detected by restriction fragment length polymorphism assay in 176 healthy subjects. RESULTS: JAK2 V617F was detected in 91 patients (59.1%): PV (91.6%), PMF (46.2%), ET (52.8%), and MPNU (66.7%). In V617F-negative MPN patients, no mutations were found in exon 12. The c.1641+179_183del5 was detected in 68.1% of V617F-negative MPN patients and 45.4% of healthy subjects (P=0.008). JAK2 V617F was closely correlated with age and leukocytosis in BCR/ABL1-negative MPN patients (P<0.05). However, c.1641+179_183del5 was not related to age, sex, or complete blood cell count parameters in V617F-negative MPN patients and healthy subjects. The c.1641+179_183del5 was associated with an increased odds ratio for MPN (odds ratio, 2.6; 95% confidences interval, 1.3-5.1; P=0.007). CONCLUSIONS: Frequencies of V617F are similar to reported results. JAK2 exon 12 mutations may be rare and c.1641+179_183del5 may influence the occurrence of MPN in Korean patients with V6 17F-negative MPN.


Assuntos
Povo Asiático/genética , Proteínas de Fusão bcr-abl/metabolismo , Variação Genética , Janus Quinase 2/genética , Transtornos Mieloproliferativos/genética , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Alelos , Substituição de Aminoácidos , Criança , Éxons , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transtornos Mieloproliferativos/diagnóstico , Razão de Chances , Polimorfismo de Fragmento de Restrição , República da Coreia , Análise de Sequência de DNA
14.
Korean J Lab Med ; 30(6): 616-23, 2010 Dec.
Artigo em Coreano | MEDLINE | ID: mdl-21157148

RESUMO

BACKGROUND: The aim of this study was to determine the yearly prevalence and genotype distribution of extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae collected over a 3-yr period in Gwangju, Korea. METHODS: Clinical isolates of E. coli and K. pneumoniae collected at Chosun University Hospital from September 15, 2005 to September 14, 2008 were evaluated. Antimicrobial susceptibility testing was performed using the Vitek II system (bioMérieux, USA) and agar dilution methods. Screening for ESBL and AmpC ß-lactamase genes was performed using PCR amplification of plasmid DNA followed by direct sequencing of the PCR products. RESULTS: The percentage of ESBL-producing isolates was 12.6% (196/1,550) for E. coli and 26.2% (294/1,121) for K. pneumoniae. The ESBL gene sequencing results showed that the most prevalent ESBL types were CTX-M (93.5%) and SHV (12.9%) in E. coli, and SHV (73.2%) and CTX-M (46.3%) in K. pneumoniae. The most common ESBL in E. coli was CTX-M-15-like, followed by CTX-M-14-like, SHV-2a-like, and SHV-12-like. The most prevalent ESBL type in K. pneumoniae was SHV-12, followed by CTX-M-14-like and CTX-M-15-like. Fifty-one percent (21/41) of ESBL-producing K. pneumoniae with ESBL types verified by sequencing also had DHA-1-like AmpC ß-lactamases. However, none of the ESBL-producing E. coli was positive in the AmpC ß-lactamase PCR analysis. CONCLUSIONS: In this study, the most common types of class A ESBLs identified were CTX-M-15-like in E. coli and SHV-12-like in K. pneumoniae.


Assuntos
Proteínas de Bactérias/genética , Escherichia coli/genética , Klebsiella pneumoniae/genética , beta-Lactamases/genética , Farmacorresistência Bacteriana/genética , Escherichia coli/isolamento & purificação , Frequência do Gene , Genótipo , Hospitais Universitários , Humanos , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , República da Coreia , Análise de Sequência de DNA
16.
Korean J Lab Med ; 29(3): 256-61, 2009 Jun.
Artigo em Coreano | MEDLINE | ID: mdl-19571625

RESUMO

A 70-yr-old woman was hospitalized with a history of dry cough. Bronchial endoscopy and transbronchial lung biopsy were performed. However, the findings of histopathology and immunohistochemistry were not sufficient to decide whether the lesion was benign or malignant, because of the presence of crush artifacts in the biopsy specimens. We performed B-cell clonality studies using BIOMED-2 multiplex PCR (InVivoScribe Technologies, USA) to detect clonal rearrangements in the immunoglobulin gene. The results of multiplex PCR showed clonal rearrangements of both kappa and lambda immunoglobulin light chain genes. The findings of immunochemistry revealed that the lesion expressed lambda light chain, but not kappa light chain. Based on the clinical, pathologic, and molecular findings, this case was diagnosed as pulmonary MALT lymphoma. We report the first case in Korea of lambda-expressing MALT lymphoma that is shown to have dual clonal rearrangements of kappa and lambda immunoglobulin light chain gene by multiplex PCR.


Assuntos
Rearranjo Gênico de Cadeia Leve de Linfócito B , Cadeias kappa de Imunoglobulina/genética , Cadeias lambda de Imunoglobulina/genética , Linfoma de Zona Marginal Tipo Células B/diagnóstico , Idoso , Feminino , Humanos , Linfoma de Zona Marginal Tipo Células B/genética , Linfoma de Zona Marginal Tipo Células B/patologia , Reação em Cadeia da Polimerase
18.
Int J Antimicrob Agents ; 32(2): 165-9, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18565738

RESUMO

An unusual class 1 integron was identified that carries an IS26-disrupted aadA1 gene cassette (designated as 'integron-IS26') in an imipenem-resistant Acinetobacter baumannii (IRAB) outbreak strain. DNA sequencing revealed that integron-IS26 contained two gene cassettes, the aac(6')-Im cassette and a peculiar aadA1 cassette that was disrupted by IS26 (disrupted aadA1 cassette). Southern blotting localised integron-IS26 to the chromosome. Nested polymerase chain reaction (PCR) was performed to define the frequency of integron-IS26 in five groups of bacteria. Nested PCR identified integron-IS26 in 19 (73.1%) of 26 clinical outbreak strains of IRAB, 10 (100%) of 10 IRAB isolated from environmental cultures, 3 (13.0%) of 23 imipenem-susceptible A. baumannii (ISAB) non-outbreak strains, 1 (3.6%) of 28 netilmicin- and tobramycin-resistant A. baumannii and none of the netilmicin- and tobramycin-resistant Pseudomonas aeruginosa. In conclusion, we have identified a novel class I integron that carries the aac(6')-Im cassette and an IS26-disrupted copy of aadA1 (integron-IS26) in most IRAB outbreak strains and in a few ISAB non-outbreak control strains. Integron-IS26 is located chromosomally.


Assuntos
Acinetobacter baumannii/genética , Aminoglicosídeos/farmacologia , Elementos de DNA Transponíveis/genética , Farmacorresistência Bacteriana/genética , Imipenem/farmacologia , Integrons/genética , Nucleotidiltransferases/genética , Infecções por Acinetobacter/epidemiologia , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/enzimologia , Antibacterianos/farmacologia , Surtos de Doenças , Humanos , Dados de Sequência Molecular , Análise de Sequência de DNA
19.
Korean J Lab Med ; 27(3): 205-9, 2007 Jun.
Artigo em Coreano | MEDLINE | ID: mdl-18094577

RESUMO

BACKGROUND: We noticed an abrupt increase in the isolation of Stenotrophomonas maltophilia from bronchoalveolar lavage (BAL) specimens collected at Chosun University Hospital. We performed surveillance cultures in order to identify the source of what appeared to be a pseudo-outbreak. METHODS: To investigate a possible nosocomial outbreak of S. maltophilia, we performed culture of 11 environmental specimens obtained from a bronchoscopy room and two bronchoscopes. Pulsed-field gel electrophoresis (PFGE) was used to examine the genetic relatedness among the strains of S. maltophilia recovered from BAL specimens of 3 patients and 1 environmental sample, as well as 9 unrelated strains of S. maltophilia as a control. RESULTS: During a 7 day-period in March 2006, S. maltophilia was isolated from the BAL specimens of 7 of 13 (54%) patients, compared to only 5 of 188 (2.6%) patients during the 6-month period prior to that period. S. maltophilia was isolated from 1 of the 11 environmental samples, which was obtained from a fiberoptic bronchoscope suction channel. All 7 patient isolates and one environmental isolate exhibited similar antibiotic susceptibility patterns. PFGE analysis of the genomic DNA from epidemic strains demonstrated an identical banding pattern, whereas each of epidemiologically unrelated strains showed a unique electrophoretic pattern. CONCLUSIONS: Apparently one of the hospital bronchoscopes became contaminated with S. maltophilia during a bronchoscopic procedure. It is likely that subsequent specimen contamination occurred because the bronchoscope had been inadequately cleaned and disinfected. The pseudo-outbreak was controlled successfully by removing the source of infection.


Assuntos
Broncoscópios/microbiologia , Surtos de Doenças , Contaminação de Equipamentos , Infecções por Bactérias Gram-Negativas/epidemiologia , Stenotrophomonas maltophilia/genética , Idoso , Idoso de 80 Anos ou mais , Líquido da Lavagem Broncoalveolar/microbiologia , Eletroforese em Gel de Campo Pulsado , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/transmissão , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Stenotrophomonas maltophilia/isolamento & purificação
20.
Korean J Lab Med ; 26(2): 86-92, 2006 Apr.
Artigo em Coreano | MEDLINE | ID: mdl-18156706

RESUMO

BACKGROUND: Group A rotavirus is a major cause of severe diarrhea in children throughout the world. For the proper management of rotavirus infections, it will be helpful to know their clinical characteristics according to the G and P genotypes of the infecting virus. METHODS: The diarrheal stool specimens from patients hospitalized in Chosun University Hospital during 2002-2003 were tested for rotavirus by Dipstick 'Eiken' Rota kit. Rotavirus antigen-positive stool specimens were analyzed for group A rotavirus by RT-PCR, and the group A-positive PCR products were genotyped for P and G types by PCR. RESULTS: Among the 119 specimens analyzed for genotypes, the predominant strain was genotype G4P[6] (51.3%), followed by G2P[4] (19.3%), G1P[8] (7.6%), G3P[8] (5.0%), and G9P[8] (4.2%). To examine the characteristics of each rotavirus genotype, a clinico-epidemiological study was performed for 100 patients whose medical records were available. The frequencies of diarrhea, vomiting, dehydration, and fever; the rates of nosocomial infection and transfer from other hospitals; and the mean severity scores were significantly different among the patients infected with different types of rotavirus. Especially, patients with G4P[6] type were more likely than those infected with other genotypes to show the following distinct features: Most patients showed milder symptoms and were neonates transferred from other obstetric hospitals and 68.4% of the cases were nosocomial infection. G4P[6] strains were isolated almost all along the year. The mean severity scores of patients infected by G4P[6], G2P[4], G1P[8], G3P[8], and G9P[8] strains were 6.8, 9.5, 8.0, 9.0, and 10.8, respectively. CONCLUSIONS: Many features of rotavirus infections including the epidemic period, rate of nosocomial infection, age and severity of symptoms were different according to the genotypes of the infecting virus.

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