耐甲氧西林金黄色葡萄球菌的分布及耐药性分析

目的分析医院耐甲氧西林金黄色葡萄球菌（MRSA）的分布及耐药情况,为临床治疗金黄色葡萄球菌医院感染提供科学依据。方法对618株金黄色葡萄球菌进行常规鉴定,用K—B法对其进行药敏试验。结果5年MRSA的平均检出率为51．9％（321／618）,MRSA感染高发主要科室为ICU、神经外科、神经内科,MRSA检出率前三位的科室为神经外科（84．1％）、ICU（76．3％）、呼吸内科（61．3％）,标本来源主要为痰液,占67．3％,检出率82．4％。MRSA对万古霉素、替考拉宁、利奈唑胺保持100％敏感,对氯霉素、米诺环素、复方新诺明等的耐药率较低,对其他药物都保持了65％以上的高耐药率。结论对重点科室监控,合理使用抗生素,严格执行无菌操作,采取有效的消毒隔离,尽量减少侵袭性操作等措施是控制并减少MRSA感染的重要环节。     

和厚朴酚抑制耐甲氧西林金黄色葡萄球菌生物被膜形成

【目的】研究和厚朴酚(HNK)抑制MRSA生物被膜(BF)形成的作用机制。【方法】使用TTC法测定了HNK对供试菌株BF的形成和成熟BF的抑制作用;刚果红平板法定性检测了HNK对PIA合成的影响;分光光度法测定了HNK对供试菌株eDNA释放量的影响;RT-PCR技术检测了HNK对供试菌株icaA、cidA以及agrA基因表达量的影响。【结果】HNK对MRSA 41573 BF的形成和成熟BF均有较强的抑制作用,其中,HNK抑制MRSA 41573 BF形成的MIC和MBC分别为10μg/mL和20μg/mL;抑制成熟BF的MIC和MBC分别为50μg/mL和100μg/mL。当用亚抑菌浓度的HNK与万古霉素联合作用后,可显著提高成熟BF对万古霉素的敏感性。HNK能显著抑制PIA的合成,且呈浓度剂量依赖。HNK能抑制供试菌株eDNA的释放量,其中1/8 MIC的HNK作用供试菌株16 h后,与对照组相比,e DNA的释放量降低了28.3%。HNK可抑制供试菌株BF形成的相关基因,其中1/2 MIC的HNK作用供试菌株16 h后,与对照相比,icaA的表达量降低了59.1%,cidA的表达量降低了56%,agrA的表达量降低了72.3%。【结论】HNK能显著抑制MRSA 41573 BF的形成,其作用机制主要是通过抑制icaA和cidA基因表达量,影响PIA和eDNA的合成,进而抑制BF的形成。此外HNK也可通过调控细菌的QS系统影响BF的形成。     

创面耐甲氧西林金黄色葡萄球菌消毒剂研究进展

创面MRSA对消毒剂产生抗性和MRSA易形成生物膜影响了创面消毒效果。本文就近年来的相关研究进展及临床策略进行了综述。     

耐甲氧西林金黄色葡萄球菌小鼠腹膜炎模型的建立

"超级细菌"耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)是诱发连续腹膜透析患者腹膜炎的常见细菌,且治疗困难。目前缺少MRSA腹膜炎动物模型。腹腔注射2×109~2×1010CFU/m L 7组不同浓度的MRSA感染小鼠,观察小鼠死亡时间,测定肝脏与脾脏细菌定植量,进行肝、脾病理分析,确定适宜的建模浓度。研究发现,小鼠感染细菌浓度最小致死剂量为每只2×109CFU,最适建模浓度为每只1.4×109CFU。结果表明建立了耐甲氧西林金黄色葡萄球菌小鼠腹膜炎模型,为MRSA致腹膜炎的致病机制研究、疫苗的研制提供实验基础。     

耐甲氧西林金黄色葡萄球菌的检测和分型方法研究进展

耐甲氧西林金黄色葡萄球菌 (Methcillin - resistantStaphylococcus aureus,MRSA )引起的院内感染 (nosocomialinfection)已经成为全世界一个越来越严重的问题。要想尽快获得 MRSA的相关信息从而采取适当的控制感染的措施 ,就必须依靠快速、可靠的检测和分型方法。由于 MRSA对甲氧西林耐药性的不断变化 ,故虽然目前存在检测和分型方法很多 ,但仍很难提供一种最优方法。在这里 ,我们对多种方法进行了比较 ,以便大家能从中选出既准确又省时且适合自己实验室使用的检测的分型方法。1　检测方法1.1　完整结构水平1.1.1琼脂平皿 2倍稀释法…     

耐甲氧西林金黄色葡萄球菌分子分型研究进展

近年来,耐甲氧西林金黄色葡萄球菌在全世界各地感染率和分离率不断提高,已成为目前院内感染的重要病原菌之一。运用有效、可靠、廉价的分子分型方法对分析耐甲氧西林金黄色葡萄球菌的流行病学特征及来源,对制定控制院感及流行的措施非常重要的。本研究概述了各种分子分型方法的原理及比较,如SCCmec分型、脉冲场凝胶电泳分型、多位点序列分型、葡萄球菌A蛋白分型和毒力因子分型等。脉冲场凝胶电泳仍然是暴发流行中MRSA分子分型的金标准,而其他分型方法更适合用于检测菌株的变异和建立国际监测。     

耐甲氧西林金黄色葡萄球菌染色体盒的研究进展

耐甲氧西林金黄色葡萄球菌（MRSA）的产生是由甲氧西林敏感的金黄色葡萄球菌（MSSA）获得外源性的SCCmec所致。MRSA菌株可以产生一种新的青霉素结合蛋白PBP2a,PBP2a降低了与β-内酰胺类抗生素的亲合力,从而对β-内酰胺类抗生素产生耐药性。PBP2a由mecA基因编码,mecA基因存在于葡萄球菌盒式染色体（Staphylococcal cassette chromosome mec,SCCmec）中,SCCmec是一种可移动的遗传元件,该元件还携带除mecA基因外的其他抗菌药物的耐药基因,造成多重耐药（Multidrug-resistance,MDR）。SCCmec目前主要分为8型,其中又分为若干亚型。SCCmec的基因型与MRSA的流行背景有关,不同地区的SCCmec基因分型分布可能不同。     

耐甲氧西林金黄色葡萄球菌分型及流行现状

【目的】研究杭州地区耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)的基因型别,探讨MRSA菌株流行变化趋势及进化特点,为该地区MRSA的进一步防治提供科学依据。【方法】对86株MRSA进行葡萄球菌盒式染色体SCCmec基因、spa基因分型,并开展多位点序列分型(Multi-locus sequence typing,MLST),与国际上MRSA的流行型别进行比较,分析进化关系。【结果】86株MRSA共发现13个spa型(以t311型为主,占48.8%;其次为t6418型,占11.6%);MLST分型共发现9个ST型(以ST5为主,占59.3%;其次为ST239,占16.3%),经e BURST软件分析它们属于4个群(Group 1、Group 6、Group 8、Group 12)和8种克隆复合体(CC1、CC5、CC630、CC20、CC59、CC88、CC239、CC573);SCCmec基因分型以SCCmecⅡ型为主,占61.6%;其次为SCCmec III型,占22%;5株社区相关性MRSA(SCCmec-Ⅳ型)。其中第一流行克隆型为SCCmec-Ⅱ-ST5-t311-CC5(占47.7%)、其次为SCCmec-III-ST239-t030/t037-CC239(占12.8%)。【结论】SCCmec-Ⅱ-ST5-t311为杭州地区当前流行菌株;CA-MRSA菌株的出现,提示MRSA菌株有由医院向社区播散的趋势;此外,对新发展了单位点变体的菌株(SCCmec-Ⅰ-ST1921-t164-CC20和SCCmec-Ⅳ-ST965-t062-CC5),应加强重视。     

耐甲氧西林金黄色葡萄球菌的分子流行病学研究

了解我院患者耐甲氧西林金黄色葡萄球菌（MRSA）的分子流行病学特点,为临床抗感染治疗提供依据。收集2007年1月~2008年9月我院分离的耐甲氧西林金黄色葡萄球菌共54株,采用PCR进行SCCmec基因分型、葡萄球菌A蛋白（SPA）分型,并检测杀白细胞毒素（PVL）基因,同时应用脉冲场凝胶电泳（PFGE）进行同源性分析。54株MRSA菌株SCCmec基因分型为SCCmecⅡ型17株,SCCmecⅢ型33株,SCCmecⅣ型2株,SCCmecⅤ型2株;SPA基因分型将28株归属为t030,9株为t002,8株为t037,5株为t570,2株为t437,t163和t796各1株;PVL毒素检测只有2株SCCmecⅣ型菌株阳性;PFGE证实院内MRSA感染主要为2种克隆株传播,同时还有其他型别出现。本院MRSA流行传播的SCCmec基因型主要以Ⅲ型占优势,同时发现有携带PVL毒素的CA-MRSA分离株流行,应引起密切关注。     

Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging contributor to biofilm-related infections. We recently reported that strains lacking sigma factor B (sigB) in the USA300 lineage of CA-MRSA are unable to develop a biofilm. Interestingly, when spent media from a USA300 sigB mutant was incubated with other S. aureus strains, biofilm formation was inhibited. Following fractionation and mass spectrometry analysis, the major anti-biofilm factor identified in the spent media was secreted thermonuclease (Nuc). Considering reports that extracellular DNA (eDNA) is an important component of the biofilm matrix, we investigated the regulation and role of Nuc in USA300. The expression of the nuc gene was increased in a sigB mutant, repressed by glucose supplementation, and was unaffected by the agr quorum-sensing system. A FRET assay for Nuc activity was developed and confirmed the regulatory results. A USA300 nuc mutant was constructed and displayed an enhanced biofilm-forming capacity, and the nuc mutant also accumulated more high molecular weight eDNA than the WT and regulatory mutant strains. Inactivation of nuc in the USA300 sigB mutant background partially repaired the sigB biofilm-negative phenotype, suggesting that nuc expression contributes to the inability of the mutant to form biofilm. To test the generality of the nuc mutant biofilm phenotypes, the mutation was introduced into other S. aureus genetic backgrounds and similar increases in biofilm formation were observed. Finally, using multiple S. aureus strains and regulatory mutants, an inverse correlation between Nuc activity and biofilm formation was demonstrated. Altogether, our findings confirm the important role for eDNA in the S. aureus biofilm matrix and indicates Nuc is a regulator of biofilm formation.     

Liposome containing cinnamon oil with antibacterial activity against methicillin-resistant Staphylococcus aureus biofilm

The global burden of bacterial disease remains high and is set against a backdrop of increasing antimicrobial resistance. There is a pressing need for highly effective and natural antibacterial agents. In this work, the anti-biofilm effect of cinnamon oil on methicillin-resistant Staphylococcus aureus was evaluated. Then, cinnamon oil was encapsulated in liposomes to enhance its chemical stability. The anti-biofilm activities of the liposome-encapsulated cinnamon oil against MRSA biofilms on stainless steel, gauze, nylon membrane and non-woven fabrics were evaluated by colony forming unit determination. Scanning electron microscopy and laser scanning confocal microscopy analyses were employed to observe the morphological changes in MRSA biofilms treated with the encapsulated cinnamon oil. As a natural and safe spice, the cinnamon oil exhibited a satisfactory antibacterial performance on MRSA and its biofilms. The application of liposomes further improves the stability of antimicrobial agents and extends the action time.     

百里香酚联合苯唑西林对耐甲氧西林金黄色葡萄球菌(MRSA)生物被膜的影响

【背景】耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)能以生物被膜的状态存在,从而产生多重耐药性和持续性感染。【目的】通过研究百里香酚和苯唑西林单用和联用对耐甲氧西林金黄色葡萄球菌生物被膜的形成抑制和清除作用,探究联合用药对MRSA生物被膜的影响,为临床联合应用抗MRSA药物提供理论依据。【方法】采用微量肉汤稀释法测定苯唑西林对MRSA标准菌株USA300的最低抑菌浓度;采用结晶紫染色法和菌落计数法评估百里香酚和苯唑西林单用和联用对USA300生物被膜形成抑制和清除作用。【结果】百里香酚和苯唑西林在亚抑菌浓度下对USA300生物被膜的形成具有一定的抑制作用。在较高浓度下,百里香酚对其24 h和72 h形成的生物被膜有良好的清除作用,而苯唑西林无清除作用。两药联用对生物被膜的抑制和清除作用进一步增强,在较低浓度下有较好的抑制和清除效果。【结论】百里香酚和苯唑西林联合用药与单独用药相比,对USA300的生物被膜的抑制和清除作用增强,两药联合有协同抗菌作用。     

两种临床标本中耐甲氧西林的金黄色葡萄球菌生物膜形成能力的检测与其耐药特征的研究

目的研究两种临床标本来源中耐甲氧西林的金黄色葡萄球菌（MRSA）生物膜形成能力和抗菌药物耐药率,并探讨二者的关系,为临床检测和治疗提供依据。方法收集温州医学院附属第二医院2009年1月至2010年12月的分离鉴定的MRSA共128株,其中脓液59株,痰液69株;采用刚果红平板检测多糖粘附素（PIA）,半定量粘附试验检测生物膜表型,PCR检测icaA基因结果用z。分割法进行统计分析。结果128株MRSA中生物膜阳性菌株为61株,其中脓液36株、痰液25株,脓液中检出生物膜的阳性率明显高于痰液中检出生物膜的阳性率（x2=7．382,P=0．005）。MRSA对所有B．内酰胺类抗生素全耐药,对万古霉素和利奈唑胺全敏感,痰液中MRSA对利福平、庆大霉素、环丙沙星的耐药率显著高于脓液中MRSA的耐药率（P〈0．05）,但是痰液中MRSA对克林霉素的耐药率要显著低于脓液中MRSA的耐药率（P〈0．05）。脓液中生物膜阳性菌株和生物膜阴性菌株对抗菌药物的耐药率差异无统计学意义（P〉0．05）,但是痰液中生物膜阳性菌株对克林霉素的耐药率高于生物膜阴性菌株的耐药率（P〈0．05）,对利福平的耐药率显著低于生物膜阴性菌株的耐药率（P〈0．05）。结论脓液中MRSA的生物膜检出率比痰液高,但是脓液中MRSA的耐药率比痰液中的耐药率要低,MRSA对抗菌药物的耐药谱和生物膜的形成并无直接的关系。     

Inhibitory effects of D-amino acids on Staphylococcus aureus biofilm development

Biofilms are communities of cells held together by a self-produced extracellular matrix typically consisting of protein, exopolysaccharide, and often DNA. A natural signal for biofilm disassembly in Bacillus subtilis is certain D-amino acids, which are incorporated into the peptidoglycan and trigger the release of the protein component of the matrix. D-amino acids also prevent biofilm formation by the related Gram-positive bacterium Staphylococcus aureus. Here we employed fluorescence microscopy and confocal laser scanning microscopy to investigate how D-amino acids prevent biofilm formation by S. aureus. We report that biofilm formation takes place in two stages, initial attachment to surfaces, resulting in small foci, and the subsequent growth of the foci into large aggregates. D-amino acids did not prevent the initial surface attachment of cells but blocked the subsequent growth of the foci into larger assemblies of cells. Using protein- and polysaccharide-specific stains, we have shown that D-amino acids inhibited the accumulation of the protein component of the matrix but had little effect on exopolysaccharide production and localization within the biofilm. We conclude that D-amino acids act in an analogous manner to prevent biofilm development in B. subtilis and S. aureus. Finally, to investigate the potential utility of D-amino acids in preventing device-related infections, we have shown that surfaces impregnated with D-amino acids were effective in preventing biofilm growth.     

The emergence and evolution of methicillin-resistant Staphylococcus aureus

Significant advances have been made in recent years in our understanding of how methicillin resistance is acquired by Staphylococcus aureus. Integration of a staphylococcal cassette chromosome mec (SCCmec) element into the chromosome converts drug-sensitive S. aureus into the notorious hospital pathogen methicilin-resistant S. aureus (MRSA), which is resistant to practically all beta-lactam antibiotics. SCCmec is a novel class of mobile genetic element that is composed of the mec gene complex encoding methicillin resistance and the ccr gene complex that encodes recombinases responsible for its mobility. These elements also carry various resistance genes for non-beta-lactam antibiotics. After acquiring an SCCmec element, MRSA undergoes several mutational events and evolves into the most difficult-to-treat pathogen in hospitals, against which all extant antibiotics including vancomycin are ineffective. Recent epidemiological data imply that MRSA has embarked on another evolutionary path as a community pathogen, as at least one novel SCCmec element seems to have been successful in converting S. aureus strains from the normal human flora into MRSA.     

Impact of biocides on biofilm formation by methicillin-resistant Staphylococcus aureus (ST239-SCCmecIII) isolates

Procedures of sterilization and disinfection are essential to ensure that medical and surgical instruments will not transmit infectious pathogens to patients. In the present paper, we tested the residual effect of these compounds on biofilm formation and its efficiency in disrupting preformed biofilms using methicillin-resistant Staphylococcus aureus (MRSA) isolates of the lineage ST239-SCCmecIII. All compounds examined, except 70% alcohol, caused a significant impairment in biofilm formation with concomitant inhibition of cell growth. Among the compounds examined, 10% povidone-iodine (PVP-I) was the only antiseptic that exhibited more than 90% reduction of both biofilm formation and dispersion. In the group of sterilants and disinfectants, a formulation containing 7% hydrogen peroxide and 0.2% peracetic acid (HP-PA), and sodium hypochlorite with 1% active chlorine (NaOCl) were equally effective.     

Resistance heterogeneity in methicillin-resistant Staphylococcus aureus

Abstract A striking feature of methicillin resistance in Staphylococcus aureus is the considerable heterogeneity of expression of resistance by cells in clonal populations: some are sensitive (or almost so), others are highly resistant, and others show intermediate resistance to the antibiotic. Subclones generally are also heterogeneous, suggesting variable inheritance or control of expression of resistance.
The degree of heterogeneity and mean resistance is influenced by environmental parameters: temperature, osmolality, pH, light, anaerobiosis, chelating agents and metal ions, and prior exposure to β-lactam antibiotics.