长沙地区临床分离金黄色葡萄球菌的耐药监测

目的了解长沙地区临床分离金黄色葡萄球菌（以下简称金葡菌）对常用抗菌药物的耐药现状,探讨金黄色葡萄球菌对甲氧西林的耐药水平。方法收集长沙地区11家医院2009年11月至2010年11月临床分离的非重复金葡菌279株,应用Vitek-2全自动微生物分析系统进行鉴定,K-B法检测金葡菌对24种药物的敏感性,产色头孢菌素试验检测β-内酰胺酶以及D试验检测诱导型克林霉素耐药。应用头孢西丁和苯唑西林纸片扩散法筛查耐甲氧西林的金葡菌（MRSA）,琼脂稀释法检测头孢西丁和苯唑西林的最低抑菌浓度（MIC）。结果在被检测的24种药物中,敏感率〉50%的药物为9种,未发现对万古霉素、替考拉宁和利奈唑胺耐药菌株;耐药率〉50%的抗菌药物有11种,其中以青霉素和氨苄西林的耐药率最高（均为97.1%）。MRSA的分离率达54.5%,且对常用的16种抗菌药物的耐药率均显著高于甲氧西林敏感金黄色葡萄球菌（MSSA）。279株金葡菌中,β-内酰胺酶阳性250株（89.6%）;红霉素耐药而克林霉素敏感或中介的30株中,D试验阳性22株（73.3%）。苯唑西林（OXA）和头孢西丁（FOX）MIC范围分别为0.125～〉256μg/mL和2～〉256μg/mL,苯唑西林的MIC50和MIC90分别为128μg/mL和256μg/mL,头孢西丁的MIC50和MIC90分别为64μg/mL和256μg/mL。结论长沙地区临床分离金葡菌对常用抗菌药物呈多重耐药;MRSA不仅分离率高,而且对甲氧西林呈高水平耐药。     

临床分离凝固酶阴性葡萄球菌细菌谱

目的了解玉溪市人民医院临床分离凝固酶阴性葡萄球菌（CNS）的菌种构成和耐药特征。方法对该院近6年来临床分离的808株CNS作回顾性分析。结果808株CNS共有16种;表皮葡萄球菌、腐生葡萄球菌和溶血葡萄球菌占CNS的77.2%;甲氧西林耐药（MRCNS）和甲氧西林敏感（MSCNS）各占63.3%和36.7%;β-内酰胺酶阳性检出率为84.5%,且MRCNS显著高于MSCNS（χ^2=50.88,P〈0.01）;药敏结果：MRCNS除万古霉素、呋喃妥因和利福平外,青霉素、氨苄西林等12种临床常用抗菌药物的耐药率在50.6%-100%,显示了高度的耐药性和多重耐药。结论CNS在临床标本中的检出范围广、种类多,以表皮葡萄球菌、腐生葡萄球菌和溶血葡萄球菌为主,MRCNS的高度耐药和多重耐药形势严重,加强CNS的细菌学监测非常必要。     

重症监护病房凝固酶阴性葡萄球菌分布及耐药性分析

目的了解重症监护病房(ICU)凝固酶阴性葡萄球菌(CNS)临床分布及耐药情况,以期指导临床合理使用抗菌药物。方法采用VITEK-2细菌鉴定及药敏分析系统,对2009年10月至2010年9月重症监护病房的患者各类标本分离的凝固酶阴性葡萄球菌进行鉴定和药敏试验。结果共检出CNS 189株,以表皮葡萄球菌和溶血葡萄球菌为主,占71.42%。耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)122株,分离率为61.5%,β-内酰胺酶检出率达100%,MRCNS对青霉素、红霉素、苯唑西林耐药率最高,分别达100%、93.44%、91.80%,对复方新诺明、喹诺酮类药的耐药率次之,对利福平、呋西地酸、替考拉宁、喹奴普汀/达福普汀等耐药率均较低;甲氧西林敏感凝固酶阴性葡萄球菌(MSCNS)67株,分离率为38.5%,β-内酰胺酶检出率为61.1%,MSCNS对大多数抗生素敏感;189株CNS中未检测到万古霉素耐药株。结论 ICU分离的CNS以表皮葡萄球菌和溶血葡萄球菌为主,MRCNS检出率高且呈多重耐药,万古霉素、呋西地酸、替考拉宁、喹奴普汀/达福普汀是治疗MRCNS感染的首选药物。     

葡萄球菌下呼吸道感染的细菌学分类及抗生素敏感性分析

目的：探讨葡萄球菌下呼吸道感染的细菌学分类情况及抗生素耐药性特点。方法：利用复星公司FOUTUNE IMS细菌鉴定药敏分析系统,凝固酶试验用试管法。结果：47株菌株分离到6种葡萄球菌,排在前3位的是施氏葡萄球菌、金黄色葡萄球菌、木糖葡萄球菌,其中金黄色葡萄球菌占29．8％。耐甲氧西林葡萄球菌（MRS）占78．7％,耐甲氧西林凝固酶阴性葡萄球菌（MRCNS）占凝固酶阴性葡萄球菌（CNS）78．8％;MRS对万古霉素敏感率为81．1％。MRS建议用药依次为万古霉素、呋喃妥因、氯霉素、四环素和喹诺酮类,未发现对万古霉素耐药的耐甲氧西林金黄色葡萄球菌（MRSA）。结论：凝固酶阴性、耐甲氧西林葡萄球菌成为下呼吸道球菌感染的主要菌株,MRS比例上升,MRS治疗首选万古霉素。     

206株凝固酶阴性葡萄球菌的临床分布及耐药性分析

目的分析凝固酶阴性葡萄球菌（CNS）的临床分布及耐药情况,指导临床合理用药。方法采用ATB-Expression细菌鉴定系统对厦门市妇幼保健院2009年1月至2009年12月分离的206株CNS进行鉴定,用K-B纸片法检测其对苯唑西林等10种抗菌药物的敏感度。结果206株CNS菌分布于8类临床标本,血液、生殖道标本分离率较高,分别为52．91％、25．24％。耐甲氧西林（MRS）的发生率分别为：模仿葡萄球菌59．09％,溶血葡萄球菌95．83％,表皮葡萄球菌80．00％,人型葡萄球菌85．71％。对青霉素、红霉素耐药率较高,分别为85％和82％以上;对左氧氟沙星、克林霉素耐药性次之;未检出对万古霉素、替考拉宁耐药菌株。结论该院分离的CNS耐药性严重,MRS发生率高达59％以上,对红霉素、克林霉素高度耐药,临床应尽量减少青霉素、红霉素等药物的经验性用药,依据药敏结果选择抗菌药物进行治疗,防止耐药菌株爆发院感流行。     

肺结核合并葡萄球菌下呼吸道感染的病原学分布与耐药性分析

目的探讨肺结核合并葡萄球菌下呼吸道感染的病原学构成和耐药特点,为临床合理使用抗菌药物提供依据。方法采用纸片扩散法对病原菌进行药敏试验,并进行耐甲氧西林葡萄球菌(MRS)检测,按CLSI 2012年标准判定药敏结果,用WHONET 5.6软件分析数据。结果 166株葡萄球菌中,主要为金黄色葡萄球菌,占58.43%;药敏试验表明3种葡萄球菌对青霉素G、苯唑西林、红霉素、克林霉素、左氧氟沙星耐药率均≥50%,对利福平、万古霉素、利奈唑胺耐药率低;调查表明本院肺结核患者耐甲氧西林葡萄球菌分离率高。结论我院肺结核合并下呼吸道感染葡萄球菌的分离率较高,且耐甲氧西林葡萄球菌所占比例高,耐药覆盖率高,呈明显的多重耐药,临床应重视葡萄球菌感染并依据药敏试验结果合理选用抗生素。     

细菌菌群的分布及耐药性分析

目的：了解本院临床分离致病菌菌群的分布及耐药情况,给临床经验用药提供可靠依据。方法：按《全国临床微生物检验操作规程》培养分离菌种,用美国BD公司的Sceptor半自动细菌鉴定仪或法国梅里埃公司的Vitek-60细菌鉴定仪进行鉴定及药物敏感实验,结果：临床分离 的致病菌中革兰阳性球菌占39．6％,革兰阴性杆菌占60．4％,前5位细菌分别为铜绿假单胞菌188株（17．0％）,大肠埃希菌143株（13．0％）,表皮葡萄球菌140株（12．7％）,肠球菌115株（10．7％）,金黄色葡萄球菌109株（9．9％）,药敏结果显示,苯唑西林耐药葡萄球菌（MRS）的耐药率明显高于苯唑西林敏感葡萄球菌109株（9．9％）,药敏结果显示,苯唑西林耐药葡萄球菌（MRS）的耐药率明显高于苯唑西林敏感葡萄球菌（MSS）,球菌中未分离出万古霉素耐药的菌株,但肠球菌中检测出有对万古霉素中介的菌株;肠杆菌科细菌对亚胺培南的耐药率最低,铜绿假单胞菌对头孢他啶,头孢哌酮,氨曲南,丁胺卡那霉素,亚胺培南的耐药率亦较低。结论：革兰阴性杆菌分离率高于革兰阳性球菌;革兰阳性球菌对万古霉素敏感率最高,肠杆菌科对亚胺培南敏感率最高,非发酵菌对亚胺培南,氨基糖甙类,头孢他啶,头孢哌酮药蓄亦较低。     

426株凝固酶阴性葡萄球菌临床分布及耐药性分析

目的:分析凝固酶阴性葡萄球菌(CNS)的分布及耐药性,为临床合理应用抗菌药物提供理论依据.方法:对我院2005年1月-2007年12月临床分离的CNS采用琼脂纸片扩散法(K-B法)检测药敏.结果:在426株CNS中共分离13种菌,其中表皮葡萄球菌、溶血葡萄球菌、人葡萄球菌分别为218株、103株、46株,各占51.17%、24.18%和10.80%;CNS对氟霉素、阿米卡星的耐药率小于25%,对万古霉素和替考拉宁的耐药率为0%;对青霉素、笨唑西林、红霉素、头孢西丁、哌拉西林、庆大霉素和环丙沙星的耐药率大于50%.从CNS中检出的耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)和甲氧西林敏感的凝固酶阴性葡萄球菌(MSCNS)分别为350株和76株,分别占82.16%和17.84%;MRCNS对多种抗菌药物耐药,与MSCNS相比差异有显著性统计学意义(p＜0.01).结论:需要加强量CNS病原学耐药性监测,了解其分布特点,及时向临床反馈公布,为临床医师合理应用抗菌药物提供依据.     

凝固酶阴性葡萄球菌致新生儿败血症临床分析

目的了解本地区凝固酶阴性葡萄球菌（CNS）致新生儿败血症的病原学及耐药性状况,用以指导临床治疗。方法回顾性分析64株CNS致新生儿败血症血培养的病原学及耐药性。结果64株CNS中以表皮葡萄球菌和溶血葡萄球菌为主;耐甲氧西林凝固酶阴性葡萄球菌（MRCNS）39株,分离率为60．9％,β-内酰胺酶检出率达100％,MRCNS对青霉素、红霉素、苯唑西林耐药率最高,对利福平、呋西地酸、替考拉宁、喹奴普汀／达福普汀等耐药率均较低;甲氧西林敏感凝固酶阴性葡萄球菌（MSCNS）25株,分离率为39．1％,13-内酰胺酶检出率为44％,MSCNS对大多数抗生素敏感;64株CNS中未检测到万古霉素耐药株。结论新生儿败血症中CNS以表皮葡萄球菌和溶血葡萄球菌为主,MRCNS检出率高且呈多重耐药,应重视对其监测与控制。     

ICU病房G^＋菌谱及其耐药性连续9年监测

目的了解我院ICU病房分离菌中G^＋菌谱及其耐药情况,为临床防治G^＋菌感染提供依据。方法对ICU病房自1998年1月至2006年12月9年间所分离到的G^＋菌及其耐药性进行回顾性调查。结果9年间共分离获得G^＋菌408株,占总的23.2%（408/1755）。由1998年至2004年,G^＋菌分离率逐年增多,2004年后又有所下降。G^＋菌以金黄色葡萄球菌（SA）为主,占65.7%,其次为溶血葡萄球菌（SH,10.8%）、表皮葡萄球菌（SE,9.5%）。MRSA占SA的89.9%,MRSE占SE的92.3%。体外药敏试验结果显示主要G^＋菌均呈多重耐药特性,只有万古霉素对G^＋具有很强抗菌活性,敏感率达100%。结论ICU病房在所分离的G^＋菌中,耐甲氧西林葡萄球菌（MRS）所占比例很高,细菌呈多重耐药特性。我们应高度重视这一严重问题并尽一切可能预防院内感染和减少多重耐药菌的产生。     

Methicillin-resistant coagulase-negative staphylococci on pig farms as a reservoir of heterogeneous staphylococcal cassette chromosome mec elements

Methicillin-resistant Staphylococcus aureus (MRSA) likely originated by acquisition of the staphylococcal cassette chromosome mec (SCCmec) from coagulase-negative staphylococci (CNS). However, it is unknown whether the same SCCmec types are present in MRSA and CNS that reside in the same niche. Here we describe a study to determine the presence of a potential mecA reservoir among CNS recovered from 10 pig farms. The 44 strains belonged to 10 different Staphylococcus species. All S. aureus strains belonged to sequence type 398 (ST398), with SCCmec types V and IVa. Type IVc, as well as types III and VI, novel subtypes of type IV, and not-typeable types, were found in CNS. S. aureus, S. epidermidis, and S. haemolyticus shared SCCmec type V. The presence of SCCmec type IVc in several staphylococcal species isolated from one pig farm is noteworthy, suggesting exchange of this SCCmec type in CNS, but the general distribution of this SCCmec type still has to be established. In conclusion, this study shows that SCCmec types among staphylococcal species on pig farms are heterogeneous. On two farms, more than one recovered staphylococcal species harbored the same SCCmec type. We conclude that staphylococci on pig farms act as a reservoir of heterogeneous SCCmec elements. These staphylococci may act as a source for transfer of SCCmec to S. aureus.     

Biomaterial-associated staphylococcal peritoneal infections in a neutropaenic mouse model

Abstract Adhesion of staphylococcal cells to polyethylene with end point-attached heparin was quantified by bioluminescence. Staphylococcus epidermidis 3380 and the slime-producing S. epidermidis RP12 adhered to the highest extent, and S. lugdunensis 2342 to the least extent. Preincubation of the polymer with dialysis fluid reduced adhesion of S. epidermidis 3380 and RP12 but enhanced that of S. aureus , and preadsorption of the surface with fibronectin decreased subsequent adhesion of S. epidermidis and S. haemolyticus strains. When staphylococci were grown in the presence of a biomaterial their ability to activate peritoneal cells was decreased. The bactericidal activity was impaired, whereas ingestion of opsonized coagulase-negative staphylococci (CNS) strains was unaffected. With S. epidermidis RP12 the presence of biomaterial did not influence either phagocytosis or bactericidal effect of peritoneal cells. After intra-peritoneal challenge with staphylococcal strains, the organ uptake of S. aureus Cowan 1 was increased in normal mice whereas immunosuppressed mice died. CNS strains increased mainly in the peritoneal cavity of immunosuppressed mice. The uptake of bacteria in liver and kidneys was increased with S. epidermidis 3380, S. lugdunensis 2343 and S. schleiferi 667-88. Generally, CNS strains persisted in the peritoneal cavity of both normal and immunosuppressed mice. These data indicate that host defense mechanisms, mainly polymorphonuclear neutrophils, fail to eliminate CNS infections in the peritoneum, and that initial adhesion to an implanted biomaterial may be of lesser importance in the peritoneal cavity than in e.g. catheter-associated infections. There are strain-specific virulence factors of bacteria, and slime producing strains evade the host defense mechanisms more efficiently than non-slime producing strains.     

凝固酶阴性葡萄球菌分布及耐药性研究

本文对１３６株ＣＮＳ进行ＡＰＩ分类,ＰＣＲ检测ＭＲＣＮＳ及纸片法药敏测定,结果表明ＣＮＳ感染以表皮葡萄球菌（占６１７６％）、溶血葡萄球菌（占２５０％）为主。ＭＲＣＮＳ已成为院内感染重要病原菌,占ＣＮＳ的７８７％。纸片法药敏结果显示ＣＮＳ对青霉素类、喹诺酮类、大环内酯类、磷霉素、氯霉素耐药较高,约为４０～７０％,有些高达８０～９０％,而对利福平、米诺环素、阿米卡星、奈替米星及某些头孢菌素类耐药率较低,对万古霉素、去甲万古霉素全部敏感,且ＭＲＣＮＳ的耐药率显著高于ＭＳＣＮＳ（ｐ＜０００１）。     

一种鉴别临床常见致病性葡萄球菌方法的建立与初步评价

目的建立一种鉴别临床常见致病性葡萄球菌的聚合酶链反应-限制性片段长度多态性（PCRRFLP）方法。方法采用经全自动微生物鉴定系统和分子生物学方法准确鉴定的金黄色葡萄球菌、表皮葡萄球菌、溶血葡萄球菌各3株,提取细菌DNA,PCR扩增tuf基因,扩增产物Alu I、Hinf I双酶切后进行琼脂糖凝胶电泳,分析不同葡萄球菌酶切后带型的差异。收集临床分离葡萄球菌142株,采用建立的PCRRFLP对其进行分类鉴别,随机选择分类鉴别的葡萄球菌各20株,PCR扩增16S r DNA,扩增产物测序,将结果与Gen Bank数据库进行比对,初步评价该方法的准确性。结果金黄色葡萄球菌,表皮葡萄球菌,溶血葡萄球菌均能扩增出长668 bp DNA片段。扩增产物经Alu I、Hinf I双酶切后电泳条带不同,金黄色葡萄球菌出现三条带（108 bp/192 bp/217 bp）,表皮葡萄球菌出现两条带（192 bp/304 bp）,溶血葡萄球菌出现两条带（192 bp/217 bp）。PCR-RFLP结果显示,142株葡萄球菌中金黄色葡葡球菌、表皮葡葡球菌和溶血葡葡球菌分别为67、29和46株。随机挑选的20株不同种葡萄球菌16S r DNA测序结果与Gen Bank数据库对应序列的相似性均〉99%,说明建立的PCR-RFLP方法能准确区分三种常见葡萄球菌。结论 PCRRFLP能准确鉴别临床常见的致病性葡萄球菌,为葡萄球菌病的分子诊断奠定了基础。     

The evaluation of lipolytic activity of strains of Staphylococcus epidermidis and Staphylococcus haemolyticus

A total of 103 isolates of CNS (66 strains of S. epidermidis and 37 strains of S. haemolyticus) were investigated. Lipolytic activity of staphylococcal strains was determined by Tryptic Soy Agar containing Tween 20 or Tween 60. The 95.4% strains of staphylococci demonstrated the lipolytic activity on Tween 20 agar and the 89.4% of strains of staphylococci degradation ester of fatty acids on Tweens 60 agar. We detected that S. epidermidis strains (respectively 95,4%, 89,4%) produced lipases more frequently than S. haemolyticus strains (respectively 72,9%, 59,4%). Studies suggest that source of isolation from clinical materials (blood, wound and pus) does not have an influence on the ability hydrolysis esters.     

Species-specific sensitivity of coagulase-negative Staphylococci to single antibiotics and their combinations

The activity of beta-lactam antibiotics (oxacillin, cloxacillin, cephalotin), vancomycin, gentamicin and rifampicin applied in vitro individually and in combination against 37 nosocomial methicillin-resistant strains of coagulase-negative staphylococci (CNS) was assessed to demonstrate the heterogeneity of this group of bacteria and estimate the chance of the efficacy of such therapy. The strains belonged to four species: Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus cohnii, Staphylococcus hominis. They originated from a hospital environment and from the skin of medical staff of the intensive care unit of a paediatric ward at a university hospital. All strains were methicillin-resistant, according to CLSI standards, but individual strains differed in MIC(ox) values. Susceptibility to other tested antibiotics was also characteristic for the species. The increased susceptibility to antibiotics in combinations, tested by calculating the fractional inhibitory concentration (FIC) index, concerned 26 out of 37 investigated strains and it was a feature of a particular species. Combinations of vancomycin and cephalotin against S. epidermidis and oxacillin with vancomycin were significant, as well as cephalotin and rifampicin in growth inhibition of multiresistant S. haemolyticus strains.     

Physico-chemical properties of staphylococci of different species in resistance to human thrombodefensins

Testing 54 strains of staphylococci (Staphylococcus aureus, S. epidermidis, S. haemolyticus, S. warneri, S. hominis, S. capitis) revealed that S. aureus in contrast to coagulase-negative staphylococci (CNS) is more resistant to bactoriocidal action of human thrombodefensins (resistance index: 60.3 vs 25.6%), less hydrophilicolipophilic balance-HLB: -0.42 vs -0.64) and less charged (x-potential: -32.4 vs -35.6 mV). In groups of staphylococci (S. aureus and CNS) correlation links of bacterial resistance to human thrombodefensins with their HLB and x-potential (r=-0.32...-0.36). By In vitro experiments, it was shown that 5 passages of staphylococci in meat-peptonic broth with human thrombodefensins (50 mkg protein/ml) lead to adaptation of bacteria followed by the formation of resistance to cationic peptides from thrombocytes, a decrease of hydrophobicity and x-potencial. The role of physico-chemical properties in providing thrombodefensin-resistance of staphylococci as a developmental factor of infectious-and-inflammatory process and persistence of bacteria was confirmed with Salmonella infection.     

Staphylococci in orthopaedic surgical wounds.

From 50 infected surgical wounds of orthopaedic patients, 43 (86%) staphylococcal strains were isolated. 34 of all these staphylococci belonged to Staphylococcus aureus species (i.e. 68 % of all isolates from surgical wounds; 79% of staphylococcal isolates); 9 were coagulase-negative staphylococci (i.e. 21% of all isolates from surgical wounds; 18% of staphylococcal isolates). Among microorganisms isolated from the wounds we also found 2 (4%) of the Enterobacteriaceae family; 2 (4%) of the Pseudomonas genus; 3 (6%) of the Streptococcus genus. Thus, orthopaedic surgical wounds were infected by staphylococci (mainly S. aureus) more frequently than by other micro-organisms. All the staphylococcal strains were screened for methicillin resistance by agar disk diffusion testing and for the presence of mecA gene responsible for methicillin resistance by PCR. 32% of the S. aureus and 33% of the S. epidermidis strains resulted methicillin resistant and mecA-positive. The data confirm the diffusion of methicillin resistant S. aureus in surgical site infections and shows that the so-called "new pathogens", i.e. S. epidermidis and other coagulase-negative staphylococci, also exhibit a frequent and hazardous methicillin-resisting ability.     

凝固酶阴性葡萄球菌耐药性及流行变迁情况的观察

通过对我院1992－1994年间的108株CNS和1997－1999年8月间的72株CNS的比较研究发现：表皮葡萄球菌的分离率仍居首位,溶血葡萄球菌和模仿葡萄球菌占的比例有所提高,耐药性的研究发现,对环丙氟哌酸和红霉素的耐药率有显著提高。     

Sensitivity to vancomycin and teicoplanin of coagulase-negative staphylococci isolated from clinical specimens

The frequency of resistance and elevated resistance to teicoplanin and vancomycin among 689 strains of coagulase-negative staphylococci isolated in one year from clinical specimens was determined. Using ATB.STAPH test, a resistance was shown mainly among strains of S. epidermidis and S. haemolyticus. The elevated resistance to teicoplanin was much more frequently observed than to vancomycin. About 27% of isolated strains of S. haemolyticus and 6.8% of S. epidermidis were classified as resistant. Among other species only single strains were recognised as resistant: one strain of S. xylosus, one of S. cohni and one of S. intermedius. 94.7% of S. epidermidis and 100% of S. haemolyticus strains classified as resistant to teicoplanin in ATB showed MIC values 14 mg/l. Moreover it was shown that 26.3% of these strains of S. epidermidis and 33.3% of S. haemolyticus had MBC of teicoplanin values equal to or higher than 32 mg/l.