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1.
Applications of fluorescent antibody for detecting capsular substances in Staphylococcus epidermidis
Y Ichiman 《The Journal of applied bacteriology》1984,56(2):311-316
A fluorescent antibody technique was developed for the determination of the type of capsule of strains of Staphylococcus epidermidis. Many mouse virulent and avirulent strain populations were investigated. Of 300 fresh isolates of Staph. epidermidis, 27 were mouse virulent strains and of these 74.1% and 25.9% were mono- and polyvalent, respectively. The frequency of capsular type antigens I, II and III, produced by the 27 virulent strains was found to be 18.5%, 88.9% and 18.5%, the majority being capsular type II. In the mouse avirulent strains, capsular type antigen production was demonstrated in 263 out of 273 strains examined and mono- and polyvalent capsular types comprised 52.3% and 44.0%, respectively. Capsular type I, II, III strains and non-typable strains occurred at frequencies of 15.0%, 95.2%, 34.9% and 3.7% respectively, the majority of mouse avirulent strains also being capsular type II. These results indicate that a majority of ordinary Staph. epidermidis produced capsular type antigens although the capability is quantitatively different according to strain. 相似文献
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Applications of fluorescent antibody for detecting capsular substances in Staphylococcus epidermidis 总被引:6,自引:3,他引:3
Y. Ichiman 《Journal of applied microbiology》1984,56(2):311-316
I chiman , Y. 1984. Applications of fluorescent antibody for detecting capsular substances in Staphylococcus epidermidis, Journal of Applied Bacteriology 56 , 311–316.
A fluorescent antibody technique was developed for the determination of the type of capsule of strains of Staphylococcus epidermidis . Many mouse virulent and avirulent strain populations were investigated. Of 300 fresh isolates of Staph. epidermidis , 27 were mouse virulent strains and of these 74.1% and 25.9% were mono- and polyvalent, respectively. The frequency of capsular type antigens I, II and III, produced by the 27 virulent strains was found to be 18.5%, 88.9% and 18.5%, the majority being capsular type II. In the mouse avirulent strains, capsular type antigen production was demonstrated in 263 out of 273 strains examined and mono- and polyvalent capsular types comprised 52.3% and 44.0%, respectively. Capsular type I, II, III strains and non-typable strains occurred at frequencies of 15.0%, 95.2%, 34.9% and 3.7% respectively, the majority of mouse avirulent strains also being capsular type II. These results indicate that a majority of ordinary Staph. epidermidis produce capsular type antigens although the capability is quantitatively different according to strain. 相似文献
A fluorescent antibody technique was developed for the determination of the type of capsule of strains of Staphylococcus epidermidis . Many mouse virulent and avirulent strain populations were investigated. Of 300 fresh isolates of Staph. epidermidis , 27 were mouse virulent strains and of these 74.1% and 25.9% were mono- and polyvalent, respectively. The frequency of capsular type antigens I, II and III, produced by the 27 virulent strains was found to be 18.5%, 88.9% and 18.5%, the majority being capsular type II. In the mouse avirulent strains, capsular type antigen production was demonstrated in 263 out of 273 strains examined and mono- and polyvalent capsular types comprised 52.3% and 44.0%, respectively. Capsular type I, II, III strains and non-typable strains occurred at frequencies of 15.0%, 95.2%, 34.9% and 3.7% respectively, the majority of mouse avirulent strains also being capsular type II. These results indicate that a majority of ordinary Staph. epidermidis produce capsular type antigens although the capability is quantitatively different according to strain. 相似文献
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The capsular polysaccharide (CPS) of Staphylococcus aureus strain Smith was labelled by growth of bacteria in the presence of radioactive N-acetylglucosamine and was separated from labelled cell wall components by affinity chromatography on wheat germ agglutinin following dissolution of the cells by lysostaphin. The products were partially characterised chemically and immunochemically. Similar labelled components were found in the culture fluid during growth. In a pulse-chase experiment, cell-bound CPS was released continuously into the culture fluid at the same rate as cell wall turnover and there was no evidence of direct excretion of CPS. 相似文献
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Abstract The capsular polysaccharide released from the bacterial surface by cell wall turnover during growth exhibited less size heterogeneity and a higher average molecular mass than the polysaccharide extracted from the cell by treatment with lysostaphin or low pH. Treatment of turnover polysaccharide, radiolabelled by growth of the bacteria in the presence of N-acetyl-[3 H]-glucosamine, with muramidase B from Chalaropsis released a low molecular weight product chromatographically identical to the peptidoglycan degradation products released from the peptidoglycan-teichoic acid complex by the same treatment. It is concluded that some or all of the capsular polysaccharide released into the culture fluid during growth is derived from peptidoglycan-linked capsular material, solubilised by cell wall turnover. 相似文献
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Genetic analysis of type 5 capsular polysaccharide expression by Staphylococcus aureus. 总被引:3,自引:1,他引:3 下载免费PDF全文
Capsules are produced by over 90% of Staphylococcus aureus strains, and approximately 25% of clinical isolates express type 5 capsular polysaccharide (CP5). We mutagenized the type 5 strain Reynolds with Tn918 to target genes involved in CP5 expression. From a capsule-deficient mutant, we cloned into a cosmid vector an approximately 26-kb EcoRI fragment containing the transposon insertion. In the absence of tetracycline selection, Tn918 was spontaneously excised, thereby resulting in a plasmid containing 9.4 kb of S. aureus DNA flanking the Tn918 insertion site. The 9.4-kb DNA fragment was used to screen a cosmid library prepared from the wild-type strain. Positive colonies were identified by colony hybridization, and a restriction map of one clone (pJCL19 with an approximately 34-kb insert) carrying the putative capsule gene region was constructed. Fragments of pJCL19 were used to probe genomic DNA digests from S. aureus strains of different capsular serotypes. Fragments on the ends of the cloned DNA hybridized to fragments of similar sizes in most of the strains examined. Blots hybridized to two fragments flanking the central region of the cloned DNA showed restriction fragment length polymorphism. A centrally located DNA fragment hybridized only to DNA from capsular types 2, 4, and 5. DNA from pJCL19 was subcloned to a shuttle vector for complementation studies. A 6.2-kb EcoRI-ClaI fragment complemented CP5 expression in a capsule-negative mutant derived by mutagenesis with ethyl methanesulfonate. These experiments provide the necessary groundwork for identifying genes involved in CP5 expression by S. aureus. 相似文献
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Staphylococcus aureus growth and type 5 capsular polysaccharide production in synthetic media. 总被引:5,自引:1,他引:4 下载免费PDF全文
The production of type 5 capsular polysaccharide by Staphylococcus aureus in synthetic media was investigated. The influence of medium components on capsular polysaccharide synthesis appeared to relate to the presence or absence of the component rather than to concentration gradient. The production of type 5 capsular polysaccharide was linked to energy availability and energy source, but not to carbohydrate concentration or carbon/nitrogen ratio. Regulation of capsular polysaccharide production by S. aureus in response to medium changes would appear to differ from that typically displayed in other organisms that produce polysaccharides. 相似文献
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以猪IgG作为捕获抗体固定金黄色葡萄球菌,修饰有万古霉素的量子点荧光微球作为"检测抗体",建立荧光酶联免疫吸附法检测金黄色葡萄球菌。文中制备了平均粒径为100 nm的量子点荧光微球并与万古霉素偶联;摸索了反应最佳盐离子浓度为0.01 mol/L,反应最佳pH为6.0。在该实验条件下,金黄色葡萄球菌的检测灵敏度为104 CFU/m L,与其他致病菌无交叉反应。以上结果表明,该方法可用于快速检测金黄色葡萄球菌,为金黄色葡萄球菌的临床监控和食品检测提供参考。 相似文献
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M Moreau J C Richards J M Fournier R A Byrd W W Karakawa W F Vann 《Carbohydrate research》1990,201(2):285-297
The Staphylococcus aureus type 5 capsular polysaccharide is composed of 2-acetamido-2-deoxy-L-fucose (1 part), 2-acetamido-2-deoxy-D-fucose (1 part), and 2-acetamido-2-deoxy-D-mannuronic acid (1 part). On the basis of methylation analysis, optical rotation, high-field one- and two-dimensional 1H- and 13C-n.m.r. experiments, and selective cleavage with 70% aqueous hydrogen fluoride, the polysaccharide was found to be a partially O-acetylated (50%) polymer of the repeating trisaccharide unit, [----4)-3-O-Ac-beta-D-ManpNAcA-(1----4)-a-L-FucpNAc-(1----3) -beta-D-FucpNAc-(1----]n. 相似文献
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Phenotypic heterogeneity and temporal expression of the capsular polysaccharide in Staphylococcus aureus 下载免费PDF全文
Shilpa E. George Tran Nguyen Tobias Geiger Christopher Weidenmaier Jean C. Lee Jan Liese Christiane Wolz 《Molecular microbiology》2015,98(6):1073-1088
Bacteria respond to ever‐changing environments through several adaptive strategies. This includes mechanisms leading to a high degree of phenotypic variability within a genetically homogeneous population. In Staphylococcus aureus, the capsular polysaccharide (CP) protects against phagocytosis, but also impedes adherence to endothelial cells and/or matrix proteins. We analysed the regulation of core biosynthesis genes (capA‐P) necessary for CP synthesis using single‐cell assays (immunofluorescence and promoter‐activity). In persistent human carriers, we found a distinct subpopulation of nasal S. aureus to be CP positive. In vitro, cap expression is also heterogeneous and strongly growth‐phase dependent. We asked whether this peculiar expression pattern (earlyOff/lateHeterogen) is orchestrated by the quorum system Agr. We show that the Agr‐driven effector molecule RNAIII promotes cap expression largely via inactivation of the repressor Rot. High NaCl, deletion of CodY or Sae also resulted in higher cap expression but did not change the earlyOFF/lateHeterogen expression pattern. Activity of the quorum system itself is largely homogenous and does not account for the observed heterogeneity of cap expression or the strictly growth phase dependent expression. Our findings are in contrast to the prevailing view that quorum sensing is the main driving force for virulence gene expression when bacterial cell densities increase. 相似文献
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Staphylococcus aureus growth and type 5 capsular polysaccharide production in synthetic media 总被引:3,自引:0,他引:3
W T Stringfellow B Dassy M Lieb J M Fournier 《Applied and environmental microbiology》1991,57(2):618-621
The production of type 5 capsular polysaccharide by Staphylococcus aureus in synthetic media was investigated. The influence of medium components on capsular polysaccharide synthesis appeared to relate to the presence or absence of the component rather than to concentration gradient. The production of type 5 capsular polysaccharide was linked to energy availability and energy source, but not to carbohydrate concentration or carbon/nitrogen ratio. Regulation of capsular polysaccharide production by S. aureus in response to medium changes would appear to differ from that typically displayed in other organisms that produce polysaccharides. 相似文献
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Five cows were inoculated intradermally with formalinized Staphylococcus aureus suspension in Freund's complete adjunvant and the development of the humoral antibody response was followed as judged by the agglutination titer of sera, at various intervals post inoculation. Highest titers were observed at 78-87 days post inoculation. Agglutinating activity was found in IgM and IgG fractions (IgG1 and IgG2) of both serum and colostrum. The agglutinating activity of colostrum was significantly higher at 12 than at 24 and 36 h, post partum. However, no such activity was detected in either normal cow serum or colostrum against S. aureus. 相似文献
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Takamitsu Miyafusa Jose?M. M. Caaveiro Yoshikazu Tanaka Martin?E. Tanner Kouhei Tsumoto 《Bioscience reports》2013,33(3)
Enzymes synthesizing the bacterial CP (capsular polysaccharide) are attractive antimicrobial targets. However, we lack critical information about the structure and mechanism of many of them. In an effort to reduce that gap, we have determined three different crystal structures of the enzyme CapE of the human pathogen Staphylococcus aureus. The structure reveals that CapE is a member of the SDR (short-chain dehydrogenase/reductase) super-family of proteins. CapE assembles in a hexameric complex stabilized by three major contact surfaces between protein subunits. Turnover of substrate and/or coenzyme induces major conformational changes at the contact interface between protein subunits, and a displacement of the substrate-binding domain with respect to the Rossmann domain. A novel dynamic element that we called the latch is essential for remodelling of the protein–protein interface. Structural and primary sequence alignment identifies a group of SDR proteins involved in polysaccharide synthesis that share the two salient features of CapE: the mobile loop (latch) and a distinctive catalytic site (MxxxK). The relevance of these structural elements was evaluated by site-directed mutagenesis. 相似文献