Isolation, molecular characteristics and disinfection of methicillin-resistant Staphylococcus aureus from ICU units in Brazil

The aim of the present study was to isolate S. aureus strains resistant to antibiotics, characterize the genotype profiles of resistance staphylococci, and evaluate the efficacy of antiseptic agents and disinfectants used in two public hospitals of Vitoria da Conquista, Bahia, Brazil. Clinical samples were obtained from ICU environments and equipment surfaces in two public hospitals in Vitoria da Conquista. Broth cultures were plated onto mannitol salt agar, and antimicrobial susceptibility testing was performed by the broth microdilution method according to CLSI. MRSA strains were submitted to PCR for detecting the mecA gene. PCR products were purified and sequenced for SCCmec type identification. Moreover, the strains were tested for efficacy of different disinfectant solutions. S. aureus were isolated from 31 and 67 sites in each hospital, respectively. Among the isolates from hospital 1, 07 (22.6%) were resistant to oxacillin while 28 (41.8%) were resistant in hospital 2. Thirty-one were positive for the mecA gene. All isolates showed SCCmec type III genotype characteristics of the Brazilian epidemic clone. In disinfectant tests, sodium hypochlorite (0.5, 1.0 and 2.0%), 2% chlorhexidine gluconate, quaternary ammonium, peracetic acid and formaldehyde were effective against the isolates tested. The strains showed higher resistance to vinegar (4% acetic acid), alcohol and glutaraldehyde. The findings of this study should assist in reducing the occurrence of nosocomial infections and therefore the morbidity, mortality and socio-economic burden caused by prolonged hospitalization.     

Comparison of methods for the detection of methicillin resistance in Staphylococcus aureus isolates from food products

AIMS: To compare several methods for detection of methicillin resistance in Staphylococcus aureus isolates from food. METHODS AND RESULTS: Two hundred S. aureus isolates from food of animal origin were screened for methicillin resistance by a PCR assay specific for the mecA gene, an oxacillin agar screen test and a cefoxitin disk diffusion test. Six out of 200 strains (3%) were found to be methicillin-resistant Staphylococcus aureus (MRSA) by PCR. The oxacillin agar screen test detected only one of the MRSA isolates (sensitivity of 16.7%) and mischaracterized three additional strains as MRSA (specificity of 98.45%). None of the MRSA strains was detected by the cefoxitin test (sensitivity of 0%), while 15 methicillin-susceptible S. aureus (MSSA) strains were misclassified as resistant (specificity of 92.3%). Fifteen MSSA strains displayed a beta-lactamase hyperproducer-like phenotype. The six MRSA (mecA-positive) strains resembled the characteristics of heteroresistant strains. CONCLUSIONS: As MRSA of animal origin may display atypical phenotypes, PCR appears to be more reliable for detection of methicillin resistance in animal strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The study stresses the need for implementing the methods of screening S. aureus from food of animal origin for methicillin resistance.     

mecA基因PCR扩增法检测耐甲氧西林金黄色葡萄球菌

目的 应用mecA基因PCR扩增法检测耐甲氧西林金黄色葡萄球菌（methicillin resistant staphylococcus aureus,MRSA）。方法 临床分离的70株金黄色葡萄球菌,应用mecA基因PCR扩增法鉴定MRSA,并与苯唑西林纸片扩散法进行比较。结果 70株金黄色葡萄球菌用PCR扩增法和纸片扩散法有6株鉴定有差异,4株。mecA基因阳性而纸片扩散法鉴定为敏感,1株mecA基因阳性纸片扩散法鉴定为临界耐药,1株mecA基因阴性却表现为苯唑西林耐药,2种方法符合率为91．43％。结论 mecA基因PCR扩增法可以准确、快速判定MRSA,特别是对隐匿型或低水平耐药菌株的检出有重要的价值。     

Sensitivity of Staphylococcus aureus strains isolated from the patients of the SP ZOZ Hospital in Nidzica to bacteriocidal agents

The aim of this study was to evaluate a frequency of isolation and antimicrobial susceptibility testing of Staphylococcus aureus strains isolated from clinical specimens obtained from patients hospitalized in different hospital wards (SP ZOZ) in Nidzica from 01. 09. 2000 to 31. 12. 2003. During over three years 716 Staphylococcus aureus strains were cultured out of 15517 clinical specimens supplied to the Bacteriological Laboratory of SP ZOZ in Nidzica. S. aureus strains were isolated from 4.6% of examined samples. Samples were collected from patients hospitalized in all wards (five wards). Analysis of susceptibility to antimicrobial agents of identified S. aureus strains was performed. Seventy strains (9.8%) were metihicillin-resistant (MRSA). One hundred twenty four strains (17.3%) revealed inducible resistance to macrolides, linkosamides and streptogramins B (MLS, mechanism). The greatest activity in vitro against clinical S. aureus strains showed glycopeptide antibiotic--vancomycin (100% of susceptible strains). Clinical S. aureus strains isolated from patients of hospital in Nidzica are in the majority susceptible to antibiotics/chemotherapeutics, except of penicillin. Percentage of methicillin-resistant strains (MRSA) is not high (<10). Nevertheless, constant monitoring of a drug susceptibility of nosocomial S. aureus strains is important, considering the necessity of control of current epidemiological and therapeutic situation.     

金黄色葡萄球菌的耐药性分析及基因分型研究

目的通过分析上海地区院内分离金黄色葡萄球菌的药敏谱型及对耐甲氧西林的金黄色葡萄球菌（MRSA）进行基因谱型的研究,了解金黄色葡萄球菌的院内流行状况。方法对临床分离出的43株金黄色葡萄球菌进行药敏试验和SCCmec基因盒的多重PCR检测,并将结果整合后用MEGA3．1软件分析其进化相关关系。结果药敏结果显示43株金葡菌对青霉素和甲氧西林的耐药率最高。甲氧西林的耐药率达到62．8％。MecA阳性菌株SCCmec的分型显示均为Ⅱ型或Ⅲ型,且所占比例相近,未见Ⅰ型和Ⅳ型。进化树分析发现了在同一医院中亲缘关系相近的菌株,为院内感染流行株。结论MecA基因介导的MRSA在分离菌株中所占比例高,存在院内感染爆发性流行。     

金黄色葡萄球菌儿童临床分离株携带PVL基因状况的分析

目的了解金黄色葡萄球菌儿童分离株携带Panton-Valentine杀白细胞素（PVL）基因的状况及感染类型。方法采用多重PCR同时检测金黄色葡萄球菌16SrRNA基因、PVL基因和mecA基因;多重PCR检测MR—SA的SCCmec基因型及亚型。结果66株金黄色葡萄球菌JL童临床分离株经多重PCR检测,其中MRSA有7株（10．6％）,MSSA有59株（89．4％）;携带PVL基因金黄色葡萄球菌有31株,总阳性率为47．O％（31／66）,其中2株为MRSA,29株为MSSA,阳性率分别为28．6％（2／7）和49。2％（29／59）。2株MRSA都属于SCCmecIV型;31株PVL基因阳性分离株有21株分离自脓液,7株分离自血液,仅1株分离自痰液。结论儿童MSSA是携带PVL基因的主要菌株,携带PVL基因的金黄色葡萄球菌主要引起化脓性感染和血流感染。     

253株金黄色葡萄球菌耐药现状分析

目的 了解医院金黄色葡萄球菌临床分布情况及其对常用抗菌药物的耐药率,为临床合理使用抗菌药物提供依据.方法 回顾分析医院2010年5月至2011年4月检出的金黄色葡萄球菌,采用VITEK-AMS全自动微生物分析仪进行菌种鉴定和药敏分析.结果 共检出金黄色葡萄球菌253株,菌株的主要来源为痰130株(51.4％)、血液39株(15.4％)、创面24株(9.5％);菌株主要科室分布前3位是神内科35株(13.8％)、ICU30( 11.8％)、脑外科26株(10.3％);其中耐甲氧西林金黄色葡萄球菌( MRSA)为165株(65.2％),MRSA对多种抗菌药物耐药率＞70.0％,MSSA为88株(34.8％),对除青霉素、红霉素外的大多数抗菌药物敏感,未发现耐万古霉素菌株.结论 MRSA检出率高,耐药现状严重,应加强对金黄色葡萄球菌耐药性的监测,并根据药敏试验结果合理使用抗菌药物.     

深圳市人民医院近五年临床分离金黄色葡萄球菌的 分布及耐药性分析

目的分析医院感染患者金黄色葡萄球菌的临床分布特征及耐药性变化,为临床治疗金黄色葡萄球菌感染提供依据。方法回顾性分析2012年1月至2016年12月我院从临床各类标本中分离获得的1 141株金黄色葡萄球菌,统计其在各类标本和病区中的分布特点,并用K-B法测定该菌对常用抗菌药物的敏感性。结果 5年中共分离出1 141株金黄色葡萄球菌,标本来源构成比最多的是伤口分泌物(43.3%)、呼吸道标本(24.0%)和血液标本(10.2%)。耐甲氧西林金黄色葡萄球菌(MRSA)共有339株,占29.7%。各年MRSA的检出数依次为53株(31.9%)、51株(26.0%)、82株(35.2%)、81株(30.3%)和72株(26.0%)。MRSA主要分离自神经外科(13.8%)、呼吸监护室(10.6%)、重症监护室(8.8%)和骨科(7.7%)。MRSA对抗菌药物的耐药性普遍高于MSSA,二者比较差异有统计学意义(P0.05)。未发现对万古霉素、利奈唑胺、替考拉宁耐药的金黄色葡萄球菌。结论 MRSA感染多发生于长期使用抗菌药物,有皮肤软组织伤口及侵入性操作的科室及患者。MRSA具有多重耐药性,应严格掌握抗菌药物的使用适应证;同时临床治疗应根据药物敏感性报告针对性地合理用药,以便及时有效地控制感染并防止耐药菌株的扩散。     

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。     

Staphylococcus aureus nosocomial infections: the role of a rapid and low-cost characterization for the establishment of a surveillance system

Continuous surveillance on resistance patterns and characterization of Staphylococcus aureus represent simple and low-cost techniques to understand and evaluate the effectiveness of infection control and antimicrobial prescribing measures. In this study we analyzed the antibiotic susceptibility and trends for S. aureus strains collected from bacteraemia cases in a five year period. Between 2004 and 2008 we noted a progressive decrease in the number of S. aureus isolates compared to all pathogens from clinical specimens and S. aureus bloodstream infections (BSI) reflected a similar trend. In particular we analyzed 185 isolates from blood cultures: 89 isolates were MSSA and 96 isolates were MRSA. Molecular SCCmec typing of these strains showed an absolute prevalence of types I and II, whereas five spa types from 96 isolates were obtained. Resistance pattern analysis allowed us to place MRSA strains into 12 antibiotypes and the major antibiotype was resistant to penicillin, gentamicin, erythromycin, clindamycin and ciprofloxacin. The predominant antibiotype among the MSSA isolates was resistant only to penicillin. In addition, 19.1% of MSSA are susceptible to all antibiotics tested. We also found a close association between antibiotyping 1 and genotyping t002/SCCmecI of MRSA strains, suggesting a nosocomial scenario dominated by a few particular clones.     

Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans

From May 2001 to April 2003, various types of specimens from cattle, pigs, and chickens were collected and examined for the presence of methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA). S. aureus was isolated and positively identified by using Gram staining, colony morphology, tests for coagulase and urease activities, and an API Staph Ident system. Among 1,913 specimens collected from the animals, 421 contained S. aureus; of these, 28 contained S. aureus resistant to concentrations of oxacillin higher than 2 micro g/ml. Isolates from 15 of the 28 specimens were positive by PCR for the mecA gene. Of the 15 mecA-positive MRSA isolates, 12 were from dairy cows and 3 were from chickens. Antimicrobial susceptibility tests of mecA-positive MRSA strains were performed by the disk diffusion method. All isolates were resistant to members of the penicillin family, such as ampicillin, oxacillin, and penicillin. All isolates were also susceptible to amikacin, vancomycin, and trimethoprim-sulfamethoxazole. To determine molecular epidemiological relatedness of these 15 animal MRSA isolates to isolates from humans, random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. The RAPD patterns of six of the isolates from animals were identical to the patterns of certain isolates from humans. The antibiotypes of the six animal isolates revealed types similar to those of the human isolates. These data suggested that the genomes of the six animal MRSA isolates were very closely related to those of some human MRSA isolates and were a possible source of human infections caused by consuming contaminated food products made from these animals.     

Incidence, antibiotic resistance and clonal relations of MRSA strains isolated from a Romanian university hospital

Our aim was to estimate the frequency and characteristics ofmethicillin-resistant Staphylococcus aureus (MRSA) strains occurring in a Romanian teaching hospital. We retrospectively studied isolates from infected or colonized patients treated at the intensive care and surgical units during January 2004-December 2005. The antibiotic susceptibility of MRSA strains and the presence of mecA gene were determined. Consecutively occurring strains isolated through a three-month period were typed using pulsed field gel electrophoresis. A total of 423 S. aureus strains were identified, methicillin-resistance was detected in 211 (49.9%) strains. Most of them were multiresistant. One of the MRSA genotypes identified by PFGE was commonly recovered from patients treated in the intensive care unit. According to our results, MRSA strains were frequently isolated pathogens in our hospital and there is an urgent need to enhance infection control efforts.     

MRSA中mecA及femB基因的检测与耐药相关性

研究femB、mecA基因在耐甲氧西林金黄色葡萄球菌(MRSA)中的表达与耐药的关系.运用PCR对MRSA的femB、mecA基因进行检测,MRSA耐药检测采用头孢西丁纸片法.40 株金黄色葡萄球菌(下简称金葡菌)通过头孢西丁纸片法,检出 30 株耐头孢西丁的菌株,通过PCR检测这 40 株金葡菌mecA基因,30 株MRSA全部为阳性, femB基因在 30 株MRSA中全部表达,而甲氧西林敏感的金黄色葡萄球菌(MSSA)的未表达.结果可见,PCR能快速准确地鉴定MRSA, mecA基因是MRSA的耐药基因,femB基因是MRSA的耐药相关基因.     

Comparative analysis of antibiotic and antimicrobial biocide susceptibility data in clinical isolates of methicillin-sensitive Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa between 1989 and 2000

AIMS: To analyse population minimum inhibitory concentrations (MICs) data from clinical strains of Staphylococcus aureus and Pseudomonas aeruginosa for changes over a 10-year period and to look for correlations between the antimicrobials tested. METHODS AND RESULTS: Data from the MIC study of 256 clinical isolates of Staph. aureus [169 methicillin-sensitive Staph. aureus (MSSA), 87 methicillin-resistant Staph. aureus (MRSA)] and 111 clinical isolates of Ps. aeruginosa against eight antimicrobial biocides and several clinically relevant antibiotics was analysed using anova, Spearman-Rho correlation and principal component analysis. Comparisons suggest that alterations in the mean susceptibility of Staph. aureus to antimicrobial biocides have occurred between 1989 and 2000, but that these changes were mirrored in MSSA and MRSA suggests that methicillin resistance has little to do with these changes. Between 1989 and 2000 a sub-population of MRSA has acquired a higher resistance to biocides, but this has not altered the antibiotic susceptibility of that group. In both Staph. aureus and Ps. aeruginosa several correlations (both positive and negative) between antibiotics and antimicrobial biocides were found. CONCLUSIONS: From the analyses of these clinical isolates it is very difficult to support a hypothesis that increased biocide resistance is a cause of increased antibiotic resistance either in Staph. aureus or in Ps. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The observation of negative correlations between antibiotics and biocides may be a useful reason for the continued use of biocides promoting hygiene in the hospital environment.     

耐甲氧西林金黄色葡萄球菌的两种新SCCmec型别及其抗药性研究

为探明本地区耐甲氧西林金黄色葡萄球菌(Staphylococcusaureus)的耐药性、流行病学分布状况及携带的葡萄球菌染色体mec盒(SCCmec)型别,用K-B琼脂扩散法、E-test和多位点PCR,对临床分离的金黄色葡萄球菌菌株进行了SCCmec分型及耐药性测定。结果发现了两种新的SCCmec型别,新1型含Ⅱ型的mecA上游特异性位点B和位于mecA内的M位点以及Ⅲ型的下游位点F,缺乏Ⅱ型上游位点C和下游位点D、G;新2型含Ⅰ、Ⅱ型的上游特异性位点A、B和两个Ⅲ型的下游位点F、H,同样缺乏位点C、D、G,可能分别为原有Ⅱ型和Ⅰ、Ⅱ型与Ⅲ型的基因重组株;且携带有新SCCmec型别的MRSA菌株,其流行病学分布特点及抗药性也与国外已报导的菌株不同,多分自门诊病人,且耐药性高,抗药谱广,值得引起高度重视和关注。     

武汉地区医院感染葡萄球菌的耐药性监测

目的了解武汉地区医院感染葡萄球菌的耐药现状。方法采用回顾性分析方法,对2003年1月到2007年12月我院分离的1373株金黄色葡萄球菌和259株表皮葡萄球菌的耐药性进行分析。药敏试验采用K—B纸片法,判断标准根据美国临床实验室标准化委员会（NCCLS）的标准。结果2003年1月到2007年12月我院分离到金黄色葡萄球菌1373株,其中耐甲氧西林的金黄色葡萄球菌（MRSA）有697株,对甲氧西林敏感株（MSSA）有587株,表皮葡萄球菌有259株,其中耐甲氧西林的表皮葡萄球菌（MRSE）有92株,对甲氧西林敏感株（MSSE）有142株。MRSA、MRSE对临床常用的抗生素几乎均耐药,只有对万古霉素和替考拉宁100％敏感;MSSA、MSSE对临床常用抗生素较敏感,但是对青霉素和红霉素耐药率均大于70％。结论武汉地区医院感染MRSA和MRSE对大部分临床常用抗生素均已高度耐药,对万古霉素和替考拉宁依然高度敏感。了解医院感染葡萄球菌的耐药状况,对临床合理选用抗生素十分重要。     

Antibiotic resistance in Staphylococci isolated from the vaginas of captive female Leontopithecus (Callitrichidae-Primates)

The purpose of this study was to provide current data on Staphylococcus species from the vaginas of clinically normal captive lion tamarins and to determine the antimicrobial susceptibility of these isolates. Samples were collected from 25 adult lion tamarins, processed to isolate Staphylococcus species, and tested for susceptibility to penicillin G, gentamicin, chloramphenicol, tetracycline, trimethoprim-sulfamethoxazole, streptomycin, ampicillin, and rifampicin. Isolates with the typical characteristics of the genus Staphylococcus were recovered from all 25 samples. Coagulase-negative species were the most common (68% of the isolates), and the most frequently isolated species (10 samples) was S. simulans. Other coagulase-negative species, including S. saprophyticus (n=5), S. epidermidis (n=1), and S. arlettae (n=1), were also recovered. Coagulase-positive Staphylococci were obtained from eight animals (six of from the S. aureus species and two from S. intermedius). Resistance to antibiotics was frequently observed, and 88% of the isolates (23 samples) showed resistance to at least one drug. Resistance to penicillin G was a common finding, and the most active antimicrobial agents were chloramphenicol and gentamicin. Coagulase-positive strains were more frequently resistant to antibiotics (79.7%, average=6.4 drugs) than coagulase-negative strains (38.2%, average=3.0 drugs). The high frequency of resistance observed in those isolates is surprising and very alarming. A detailed history of the use of antimicrobial drugs in these subjects did not reveal any previous exposure to any of the tested antibiotics that could justify the observed resistance rate.