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1.
Yum JH Kim S Lee H Yong D Lee K Cho SN Chong Y 《Journal of Korean medical science》2005,20(6):961-965
Respiratory isolates of Klebsiella pneumoniae in Korea during 2002-2003 were studied to determine the prevalence and types of extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases (PABLs). ESBL-production was tested by double-disk synergy, and genotypes of beta-lactamases were determined by PCR and sequencing. ESBLs were detected in 28.4% of 373 isolates, and the most prevalent types were SHV-12 (63 isolates) and CTX-M-14 (9 isolates). Forty of 75 ESBL-producers (53.5%) also had PABLs: 21 isolates with CMY-2-like, 17 with DHA-1-like. Pulsed-field gel electrophoresis showed 19 types and 25 of 74 isolates had an identical pattern, indicating nosocomial spread. Dissemination of ESBL- and PABL-producing K. pneumoniae strains in Korea is a particular concern, as it limits the choice of antimicrobial agents for treatment of infections. 相似文献
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Persistent increase of matrix metalloproteinases in cerebrospinal fluid of tuberculous meningitis 总被引:7,自引:0,他引:7
Lee KY Kim EH Yang WS Ryu H Cho SN Lee BI Heo JH 《Journal of the neurological sciences》2004,220(1-2):73-78
Matrix metalloproteinase (MMP)-2 and MMP-9 were analyzed by gelatin zymography and an enzyme-linked immunosorbent assay (ELISA) in a cerebrospinal fluid (CSF) from patients with tuberculous meningitis (n=24), acute aseptic meningitis (n=23) and the control (n=10). The MMP-2 and MMP-9 levels were significantly higher in the samples from the tuberculous meningitis patients than those from either the aseptic meningitis patients or the controls. In tuberculous meningitis, the patients with late neurologic complications had higher MMP-2 and MMP-9 levels than those without. The persistent increase in the MMP-2 and MMP-9 levels was associated with the development of complications following tuberculous meningitis. Inhibiting the MMPs may be an effective strategy for preventing or reducing the complications in tuberculous meningitis. 相似文献
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Effect of vaccination with refined components of the organism on infection of mice with Mycobacterium leprae
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Ngamying M Sawanpanyalert P Butraporn R Nikasri J Cho SN Levy L Brennan PJ 《Infection and immunity》2003,71(3):1596-1598
Only native products of Mycobacterium leprae, whether cell wall, cytosol, or membrane derived, can confer protective immunity against challenge in the mouse footpad. Previously, recombinant proteins were shown to be ineffective. The cell wall skeleton-the mycolyl-arabinogalactan-peptidoglycan complex-devoid of proteins is not protective. 相似文献
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Comparative analysis of effects of cytokine gene adjuvants on DNA vaccination against Mycobacterium tuberculosis heat shock protein 65 总被引:11,自引:0,他引:11
Baek KM Ko SY Lee M Lee JS Kim JO Ko HJ Lee JW Lee SH Cho SN Kang CY 《Vaccine》2003,21(25-26):3684-3689
DNA-based vaccines generate potent cellular immunity as well as humoral immunity. It seems evident that cytokines play a crucial role in generation of effector T cell subsets and in determining the magnitude of the response by DNA vaccines. In this study, we compared the effects of several TH1 cytokine genes as adjuvant in DNA vaccination using mycobacterial Hsp65 as a model antigen. Our results demonstrated that although the overall immune response to Hsp65 was enhanced by co-injection of Hsp65 DNA with cytokine genes, each cytokine gene was shown to affect different immune response elements. Co-injection of Hsp65 DNA with IL-12 or GM-CSF led to an increase in IFN-gamma production and represented potent protections against Mycobacterium tuberculosis challenge, while that with Eta-1, IL-12 or IL-18 gene led to an elevated IgG2a/IgG1 ratio. Interestingly, co-administration of Flt3L gene was shown to enhance the Ag-specific CTL response. These results show that the direction and magnitude of immune response in DNA vaccination against Hsp65 of M. tuberculosis could be modulated in different ways by co-injection of an appropriate cytokine gene as adjuvant. 相似文献
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Miyako Kimura Rama Murthy Sakamuri Nathan A. Groathouse Becky L. Rivoire David Gingrich Susan Krueger-Koplin Sang-Nae Cho Patrick J. Brennan Varalakshmi Vissa 《Journal of clinical microbiology》2009,47(6):1757-1766
Mycobacterium leprae is the noncultivable pathogen of leprosy. Since the genome sequence of an isolate of M. leprae has become available, multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) has been explored as a tool for strain typing and identification of chains of transmission of leprosy. In order to discover VNTRs and develop methods transferable to clinical samples, MLVA was applied to a global collection of M. leprae isolates derived from leprosy patients and propagated in armadillo hosts. PCR amplification, agarose gel electrophoresis, and sequencing methods were applied to DNA extracts from these infected armadillo tissues (n = 21). We identified polymorphisms in 15 out of 25 short-tandem-repeat (STR) loci previously selected by in silico analyses of the M. leprae genome. We then developed multiplex PCR for amplification of these 15 loci in four separate PCRs suitable for fluorescent fragment length analysis and demonstrated STR profiles highly concordant with those from the sequencing methods. Subsequently, we extended this method to DNA extracts from human clinical specimens, such as skin biopsy specimens (n = 30). With these techniques, mapping of multiple loci and differentiation of genotypes have been possible using total DNA extracts from limited amounts of clinical samples at a reduced cost and with less time. These practical methods are therefore available and applicable to answer focused epidemiological questions and to allow monitoring of the transmission of M. leprae in different countries where leprosy is endemic.The causative pathogen of leprosy is Mycobacterium leprae. A continued incidence, defying global campaigns to eliminate leprosy even after years of rigorous case finding and the availability of multidrug therapy regimens (28, 29, 30, 31), is attributed to subclinical human and environmental reservoirs of the pathogen (1, 8, 13). In recent years, molecular strain-typing methodologies have complemented conventional infectious disease epidemiology. With the publication in 2001 of the complete genome sequence of an isolate from Tamil Nadu, India, called the TN strain (4), selection of potential polymorphic genomic markers for strain typing was feasible. The first genetic markers that showed polymorphism were short tandem repeats (STRs) in the M. leprae genome. One was a 6-bp intragenic sequence in the rpoT gene, and the second, a trinucleotide (TTC) repeat element upstream of a pseudogene (17, 23). These sequences exhibit variable numbers of tandem repeats (VNTRs) when sequenced in different isolates. Based on these observations, we short-listed 44 loci (including the rpoT and TTC loci) by in silico analyses of the M. leprae genome and accomplished the screening of 11 STR loci, of which 9 were polymorphic when tested in a small panel of four human isolates derived from passage through armadillos (6). Five were minisatellites (6- to 50-bp repeat units), and four were microsatellites (1- to 5-bp repeat units). Since then, others have also shown that VNTR loci exist in M. leprae isolates (25, 33, 34). Three single-nucleotide polymorphisms have also been discovered by comparing sequences of a limited number of strains (20).The goal of our work has been to discover and apply DNA variation among M. leprae isolates to identify sources and chains of transmission of leprosy in regions of endemicity. There are, however, physiological and practical issues relevant to strain typing of M. leprae in the clinical setting, such as the long incubation period and low transmissibility of leprosy and the requirement for clinical specimens such as slit skin smears and skin biopsy specimens from leprosy patients due to the inability of M. leprae to grow in culture. During the course of the last 4 years, field studies in which STR mapping was implemented have been reported. Matsuoka et al. (16) applied the microsatellite locus (TTC)21 to type M. leprae strains obtained from nasal swabs and slit skin smears from patients grouped by village, dwelling, or household in Indonesia, while Young et al. (33) combined (AT)15, (GTA)9, and (TTC)21 VNTR loci for the identification of short- and long-range M. leprae transmission chains in areas within and surrounding the city of Hyderabad, India. Monot et al. mapped five M. leprae STR loci in patients from Mali, Africa (19). The results from those studies demonstrated heterogeneity in prevalent haplotypes, indicating that genotype mapping with a small panel of one to five microsatellite VNTR loci was insufficient to discern strain relatedness. However, within an intrafamilial case, three markers were congruent (33). The authors of those studies concluded that in these areas of endemicity, multiple rather than single dominant isolates are found and that additional genomic markers are necessary for strain typing.For these reasons, assays for the amplification and differentiation of multiple genomic loci are needed. When these requirements have been met, it becomes possible to undertake systematic strain-typing studies that include suitable sampling strategies and conventional epidemiology methods for monitoring transmission and detecting clusters of cases. In light of these laboratory, field, and clinical issues, we further explored multiple-locus VNTR analysis (MLVA) techniques. In this paper, we report the development and testing of multiplex-PCR methods for MLVA for reference armadillo-derived M. leprae isolates and clinical materials and address allelic properties of individual loci and the reproducibility and feasibility of the techniques. In a future paper, we will apply and extend these methods to the data from population-based studies in Cebu, the Philippines (R. M. Sakamuri, M. Kimura, W. Li, K. Madanahally, H.-C. Kim, H. Lee, M. Balagon, R. Gelber, W. C. Black, S.-N. Cho, P. J. Brennan, and V. Vissa, submitted for publication). 相似文献
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Yoon JH Nam JS Kim KJ Choi Y Lee H Cho SN Ro YT 《Diagnostic microbiology and infectious disease》2012,72(1):52-61
We investigated the causal relationship between genotype and phenotype of drug-resistant Mycobacterium tuberculosis isolates obtained from patients with pulmonary tuberculosis (TB) in Korea. Of 80 isolates tested, 17, 20, 1, and 7 isolates were mono-resistant to ethambutol (EMB), isoniazid (INH), pyrazinamide (PZA), and rifampicin (RFP), respectively, and 31 isolates (38.8%) were multidrug-resistant (MDR). Sequencing analysis showed that 78% (32/41) of RFP-resistant strains had mutations in the rifampicin resistance-determining region (RRDR) of rpoB, and the mutation at rpoB531 (59.4%) was most abundant. In 52 INH-resistant strains, mutations were found mostly at C-15T (n = 21, 40.4%) in the inhA promoter region as well as at katG315 (n = 12, 23.1%). Mutations at embB306 were mostly found in 26.7% (12/45) of EMB-resistant isolates. New mutations found here in MDR isolates include rpoB523 (Gly523Glu) and embB319 (Tyr319Ser). Consequently, mutations in the rpoB531, C-15T in the inhA promoter region, embB306, and katG315 would be a useful marker for rapid detection of MDR M. tuberculosis isolates in Korea. 相似文献
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Park SK Hong S Eum SY Lee IH Shin DO Cho JE Cho S Cho SN 《Tuberculosis (Edinburgh, Scotland)》2011,91(4):300-307
The immune responses of multidrug-resistant tuberculosis (MDR-TB) patients undergoing lung resection surgery were investigated in order to understand the mechanism of strong immune suppression in MDR-TB. We examined changes in cell-mediated immune response (CMI) of a total of sixteen MDR-TB patients, three of them extensively drug-resistant tuberculosis (XDR-TB) patients, after the removal of the heavily diseased lung section. The IFN-γ response to Mycobacterium tuberculosis culture filtrate proteins (Mtb-CFP), one of the most important CMI to defend TB, showed a statistically significant elevation in 2-4 months after operation when compared to the preoperative CMI in patients who were converted into AFB negative and cured in two years' follow-up, suggesting that the recovery of CMI may be one of the key factors in the successful treatment of MDR-TB. Interestingly, IL-10 response to Mtb-CFP was also elevated in 2-4 months after surgery in cured patients although both proliferative response and PBMC composition were not significantly changed. Infection with first- or second-line drugs resistant Mtb reduces the efficiency of chemotherapeutic treatment of MDR-TB to about 50%. Thus, this study suggests that chemotherapeutic treatment of MDR-TB may be more effective when combined with accompanying therapy that increases CMI, includes lung resection surgery. 相似文献