中重度AECOPD感染铜绿假单胞菌的耐药表型及基因型分析

目的 探讨中重度慢性阻塞性肺疾病急性加重(AECOPD)感染铜绿假单胞菌耐药现状.方法 AECOPD患者105例,经呼吸道分泌物分离出铜绿假单胞菌21株,体外采用K-B法测定对14种抗生素的药敏结果.用PCR法检测14种耐药基因,分析铜绿假单胞菌的耐药表型和基因型.结果 21株铜绿假单胞菌对抗生素的耐药率:头孢曲松80.9％,头孢他啶76.2％,头孢吡肟66.7％,亚胺培南14.4％.21株铜绿假单胞菌中,blaTEM、blaSHV、blaOXA-10、blaPER、blaDHA阳性率分别为28.6％、21.4％、14.2％、42.8％、7.1％,而blaVEB、blaCTX-M、blaIMP、blaVIM、blaGIM、blaSPM均为阴性,oprD2基因缺失率为7.2％,ant(3″)Ⅰ,aac(6′)-Ⅱ阳性率分别为35.7％,36.8％.结论 中重度AECOPD分离的铜绿假单胞菌携带多种耐药基因,耐药基因主要与β-内酰胺类基因blaTEM、blaSHV、blaPER、blaOXA-10有关.     

慢性阻塞性肺病合并铜绿假单胞菌感染32例

近年来铜绿假单胞菌 (ｐｓｅｕｄｏｍｏｎｅｓａｅｒｕｇｉｎｏｓａ ,简称ＰＡ)肺部感染逐年增加 ,尤其高龄人群 ,由于机体免疫力低下 ,ＰＡ成为呼吸道感染的重要致病菌 ,且治疗较困难、预后差。我院 1996年 1月～ 1999年 5月共收治此类患者 3 2例 ,报道如下。1　临床资料1.1　一般资料　男 2 3例 ,女 9例。年龄 5 4～ 88岁 ,平均66.7岁。全部病例均为慢性阻塞性肺病 (ＣＯＰＤ) ,曾反复使用多种抗生素者 2 7例 ( 84.4% ) ,应用皮质激素者 12例 ( 3 7.5 % )。本组临床均有肺部感染的症状及体征 ,符合ＣＯＰＤ的诊断 ,胸部Ｘ线有炎症阴影 ,痰…     

老年COPD患者铜绿假单胞菌感染耐药分析

目的:分析老年COPD患者急性加重期铜绿假单胞菌感染的耐药状态.方法:采用常规方法分离、鉴定病原菌,选用13种常用抗生素对痰液中分离的铜绿假单胞菌进行体外MIC药敏试验.结果:铜绿假单胞菌对亚胺培南的耐药率最低,为8.5%,其次是头孢哌酮/舒巴坦和头孢他啶,均为10.6%;耐药率最高的是头孢噻肟(91.5%)和头孢曲松(83.0%).结论:铜绿假单胞菌是老年COPD患者急性加重期下呼吸道感染的主要致病菌,在临床治疗中应根据药敏结果选用抗生素.     

铜绿假单胞菌慢性肺部感染合并免疫低下大鼠模型的建立

目的：建立铜绿假单胞菌（Pseudomonas aeruginosa,PA）慢性肺部感染合并免疫低下大鼠模型,为相关模型建立与研究提供依据。方法：将实验大鼠分为空白对照组（A组）、肺部感染组（B组）、免疫低下组（C组）、肺部感染合并免疫低下组（D组）。以40 mg·kg^-1的剂量给予C、D 2组大鼠腹腔注射环磷酰胺（cyclophosphamide,CTX）,连续给药3 d,造模成功后以相同剂量每周注射一次;运用气管插管法,构建B、D组大鼠慢性肺部感染。在模型建立期间,以固定时间对大鼠肺载菌量、肺部病理学变化、免疫学指标变化、T细胞亚群等指标进行考察。结果：HE染色结果显示,B、D组大鼠在造模成功后肺部产生病理学病变,并有较明显的中性粒细胞聚集等现象。与A组相比,B组与D组大鼠外周血中性粒细胞有不同程度的增高,C组与D组白细胞数目有所下降,实验组大鼠CD₄⁺/CD₈⁺有显著增高。结论：通过该实验方法,成功建立PA慢性肺部感染合并免疫低下大鼠模型,成功率高、死亡率低,适于采取该方法进行相关研究。     

233例感染铜绿假单胞菌的老年患者耐药性分析

目的了解感染铜绿假单胞菌老年患者的耐药性特征,为临床预防及治疗提供理论依据。方法收集我院2013年1月至2014年7月感染铜绿假单胞菌的老年患者病例233例,采用Kirby-Bauer纸片扩散法和CLSI 2013年版判断标准进行耐药性监测,并用SPSS 19.0统计软件进行数据分析。结果阿米卡星耐药率最低,其他耐药率低于60%的包括庆大霉素、妥布霉素、复方新诺明、罗米沙星、左氧氟沙星、环丙沙星、美罗培南、亚胺培南、头孢吡肟、哌拉西林、哌拉西林/他唑巴坦、头孢他啶。结论老年患者中分离的铜绿假单胞菌呈现出多重耐药且耐药率不断上升的特点,建议根据药敏结果选择抗菌药物并联合用药。     

慢性阻塞性肺疾病合并铜绿假单胞菌感染的诊治分析

目的通过研究分析慢性阻塞性肺疾病合并铜绿假单胞感染的状况和耐药性,探讨治疗方案及早期预防措施。方法随机选取60例2015年3月到我院就诊的慢性阻塞性肺疾病合并铜绿假单胞菌感染患者,通过对这60例慢性阻塞性肺疾病患者采集痰标本,再将痰标本送去检验进行培养,最后分离出铜绿单胞菌进行药物测试和统计分析。结果通过鉴定和药物测试,这60株铜绿假单胞菌对阿莫西林的耐药性最高,达到了86.32%;对阿米卡星、西司他丁的耐药性较低,仅14.28%和17.43%;对其余的环丙沙星、头孢曲松等药物的耐药性在20%~60%。同时经过分析,大多数抗菌药物的耐药率逐年增高。结论铜绿假单胞菌对多种药物的耐药性较高,并且耐药性也逐年增高,这是慢性阻塞性肺疾病合并铜绿假单胞菌感染患者治疗时的重难点。医学工作者要重视对铜绿假单胞菌对多种药物的耐药性研究,合理使用抗菌药物,并加强慢性阻塞性肺疾病合并铜绿假单胞菌感染患者的感染控制。     

1例铜绿假单胞菌合并烟曲霉菌肺部感染患者的个体化治疗

摘要：本文报道临床药师通过全程参与1例铜绿假单胞菌合并烟曲霉菌肺部感染的临床治疗,通过患者进本情况与病情变化及时调整药物,并为患者制定个体化的抗感染治疗方案,取得了满意的临床疗效。在临床治疗中体现了临床药师的价值,提高了治疗的安全性和有效性。     

1例马尔尼菲篮状菌合并铜绿假单胞菌重症肺炎患者抗感染治疗的病例分析

1例70岁老年男性患者因双肺炎症入院,临床表现为咳嗽、咯痰、气喘、发热;胸部CT影像学表现为疾病初期以云絮状、斑片影为主,恢复期肺部实变影吸收并出现\"磨玻璃\"样改变;痰培养提示铜绿假单胞菌和酵母样真菌,予以比阿培南、伏立康唑等药物抗感染,治疗显效;后肺泡灌洗液病原体宏基因测序检出马尔尼菲篮状菌。临床上马尔尼菲篮状菌感染少见,其临床症状和影像学表现缺少特异性,从临床标本分离培养出具有双相型的马尔尼菲篮状菌是确诊的金标准,但培养耗时长,因而该患者抗真菌用药与明确病原菌存在较长时间差。患者在治疗过程中出现了肝功能异常,临床药师分析伏立康唑所致的肝功能异常可能性大,但转氨酶未达正常值的5倍,肝功能异常的临床症状不明显,可在监测肝胆指标的情况下继续使用伏立康唑,必要时给予还原型谷胱甘肽降酶进一步护肝处理。     

In vitro activities of ceftobiprole combined with amikacin or levofloxacin against Pseudomonas aeruginosa: evidence of a synergistic effect using time-kill methodology

Ceftobiprole is an investigational intravenous broad-spectrum cephalosporin with in vitro activity against Gram-positive and Gram-negative pathogens, including meticillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Pseudomonas aeruginosa is a frequent nosocomial pathogen, increasingly associated with complicated skin and skin-structure infections. Combination antimicrobial therapy is recommended as empirical therapy for serious infections where P. aeruginosa is suspected. Therefore, in this study the interaction of ceftobiprole with two other antipseudomonal agents (amikacin and levofloxacin) was investigated. Time-kill studies were performed for each single agent and for the combination of ceftobiprole 4 mg/L with either amikacin or levofloxacin at 0.5×, 1× and 2× the minimum inhibitory concentration. Five clinical isolates of P. aeruginosa as well as the P. aeruginosa ATCC 27853 reference strain were tested at initial inocula of 5 × 10⁵ colony-forming units (CFU)/mL (low inoculum) or 5 × 10⁷ CFU/mL (high inoculum). Synergy was defined as a decrease of ≥2 log₁₀ CFU/mL with the combination compared with the most active single drug at 6 h and 24 h. At low inoculum with ceftobiprole as a single agent, viable counts were decreased by 1.5-2 log₁₀ at 6 h. Addition of either amikacin or levofloxacin resulted in synergistic bactericidal activity at 24 h. At high inoculum the combination of ceftobiprole with amikacin or levofloxacin demonstrated synergism in one of three and three of five strains, respectively. This study demonstrated that the combination of ceftobiprole at a clinically achievable concentration of 4 mg/L with amikacin or levofloxacin exhibited synergistic activity against P. aeruginosa. There was no evidence of antagonism for either combination.     

High burden of antimicrobial resistance in Asia

Pseudomonas aeruginosa is a nosocomial and community-acquired pathogen associated with considerable patient morbidity and mortality. Multidrug resistance in P. aeruginosa is a concern owing to the limited therapeutic options available to treat infections due to this organism. In this study, rates of antimicrobial resistance of P. aeruginosa isolates collected by The Surveillance Network Database-USA (Eurofins Medinet, Chantilly, VA) from 1997 to 2009 were examined. The patient population and specimens were stratified according to patient setting and age as well as specimen source. Multidrug resistance was defined as resistance to three or more of the following antimicrobial agents: aztreonam; cefepime; ciprofloxacin; imipenem; gentamicin; and piperacillin/tazobactam (TZP). A total of 924 740 P. aeruginosa isolates were examined in this study. Changes in resistance rates to individual antimicrobial agents were <5% for all agents except ciprofloxacin. There was a statistically significant decreasing rate of multidrug-resistant P. aeruginosa to four, five and six antimicrobial agents. For isolates resistant to imipenem, aztreonam and gentamicin, ciprofloxacin had the highest cross-resistance rates. The greatest coverage against P. aeruginosa was by the combination of TZP plus amikacin (94%) followed by aztreonam plus amikacin (90%). Pseudomonas aeruginosa resistance rates remained steady or minimally declined to all antimicrobials from 1997 to 2009. Amongst the β-lactams, TZP has the greatest activity against P. aeruginosa.     

临床药师参与1例小儿烫伤后感染治疗方案的制定

目的 探讨临床药师在重症感染病例救治中的作用。方法 临床药师通过参与1例小儿烫伤后感染治疗方案的会诊工作, 对抗感染药物的选择提出具体意见, 为患儿制定个体化的给药方案。结果 重症感染的患儿得到及时控制和治疗。结论 临床药师参与临床治疗, 有利于提高临床治疗水平, 促进合理用药。     

Antimicrobial effect of resveratrol on dermatophytes and bacterial pathogens of the skin

The phytoalexin resveratrol is commonly found in food and drinks, including red wine, grapes, and peanuts. Many studies have shown that this compound has anti-inflammatory properties, and it has been ascribed as having health benefits that help to prevent cancer and coronary heart disease. A treatment that combines antimicrobial and anti-inflammatory actions may be desirable for alleviating many skin conditions that range in severity. Therefore, we evaluated the antimicrobial activity of resveratrol against bacteria and dermatophytes that are major etiologic agents of human skin infections. Using the broth microdilution protocol of the National Committee for Clinical Laboratory Standards (NCCLS) M7-A5, growth of the bacterial species Staphylococcus aureus, Enterococcus faecalis, and Pseudomonas aeruginosa was inhibited at 171-342 microg/mL of resveratrol in the solvent dimethyl sulfoxide. Using the NCCLS protocol M38-P, activity against the fungal species Trichophyton mentagrophytes, Trichophyton tonsurans, Trichophyton rubrum, Epidermophyton floccosum, and Microsporum gypseum was also tested. The growth of dermatophytes was inhibited at 25-50 microg/mL of resveratrol. Thus, this study indicates a novel application for resveratrol, a molecule of plant defense, to combat human fungal pathogens. Resveratrol and its analogs may have wide application to skin conditions that afflict a significant portion of our population, and may also have promising clinical potentials in diabetic wounds.     

Prevalence of efflux-mediated ciprofloxacin and levofloxacin resistance in recent clinical isolates of Pseudomonas aeruginosa and its reversal by the efflux pump inhibitors 1-(1-naphthylmethyl)-piperazine and phenylalanine-arginine-β-naphthylamide

To assess the prevalence of efflux-driven fluoroquinolone (FQ) resistance in recent clinical isolates of Pseudomonas aeruginosa, a worrisome and often hospital-acquired pathogen, 115 unique strains were collected over a 5-month period, of which 27 and 33 had decreased susceptibility to ciprofloxacin (CIP) and levofloxacin (LVX), respectively. The MIC₅₀ (minimum inhibitory concentration for 50% of the organisms) was 16 μg/mL for both FQs. The efflux pump inhibitors (EPIs) phenylalanine-arginine-β-naphthylamide (PAβN) and 1-(1-naphthylmethyl)-piperazine (NMP) were then used to evaluate their efficacy in reducing CIP and LVX MICs. NMP did not significantly modify CIP MICs, whilst PAβN resulted in MIC₅₀ values of 2 μg/mL and 0.125 μg/mL for CIP and LVX, respectively. With the addition of PAβN, susceptibility to CIP and LVX was recovered in 6 (22.2%) and 31 (93.9%) strains, respectively. The best combination to reverse FQ resistance in this set of strains was LVX with PAβN. The results of this study show that the effect of an EPI is not only dependent on the species on which it is used but also on the molecule associated with it. Therefore, the design of an EPI equally efficient on all resistance-nodulation-cell division (RND) efflux pumps appears to be difficult and, from a practical point of view, if an EPI is developed for clinical use, the efficiency of its combination with a definite molecule should be assessed carefully against a wide range of clinical isolates to evaluate the real benefit of this combination.     

Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa

Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. To reduce the selection pressure for resistance, it is important to determine the antibiotic susceptibility pattern of bacteria so that hospital patients can be treated with more narrow-spectrum and target-specific antibiotics. This study describes the development of a technique for detecting point muations in the fluoroquinolone resistance-determining region of the gyrA and parC genes as well as the efflux regulatory genes mexR, mexZ and mexOZ that are associated with fluoroquinolone and aminoglycoside resistance. The assay is based on a short DNA sequencing method using multiplex-fast polymerase chain reaction (PCR) and Pyrosequencing™ for amplification and sequencing of the selected genes. Fifty-nine clinical isolates of P. aeruginosa were examined for mutations in the abovementioned genes. Mutations related to antibiotic resistance were detected in codons 83 and 87 of gyrA and codon 126 of the mexR regulatory gene. Results of this study suggest Pyrosequencing™ as a substitute for traditional methods as it provides a rapid and reliable technique for determining the antibiotic resistance pattern of a given bacterial strain in <1 h.     

Dynamics of glutathione-S-transferases in Mytilus galloprovincialis exposed to toxic Microcystis aeruginosa cells, extracts and pure toxins

Molluscs and especially bivalves are able to accumulate dinoflagelates, diatoms and cyanobacteria toxins, and, being vectors for these toxins, transfer them along food chains. The data obtained from laboratory experiments showed that bivalve molluscs are resistant to cyanobacteria toxins. In this work, we wanted to test if Mytilus galloprovincialis organs react to microcystins and other cyanobacteria compounds by inducing or decreasing its GST activity. Acclimated mussels M. galloprovincialis were exposed to the toxic Microcystis aeruginosa M13 strain. Exposure of mussels to toxins was done in three ways: living Microcystis cells, crude Microcystis extracts and pure toxins. The measurement of soluble and microsomal GST activity in the different mussel organs was done by using the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 2,4-dichloro-1-nitrobenzene (DCNB). Analysis of the GST activity of the control mussels using CDNB as a substrate showed that cytosolic activity is much more significant than microsomal. Intact M. aeruginosa cells did not induce any significant response from the mussels, showing that these animals are quite resistant to the cyanobacteria if they are intact. On the other hand, cell extracts caused an important effect in the gut, in the gills and in the labial palps, although in different ways. There was an increase in GST activity in the gut and gills of mussels exposed to Microcystis extracts, showing a response of this detoxication pathway, but in the labial palps a severe reduction in GST activity occurred. Pure MC LR+YR induced an increase in GST activity in all organs but the labial palps. The results showed that other substances apart from microcystins may cause stress to mussels and affect detoxication enzymes such as GST.     

铜绿假单胞菌合并按蚊伊丽莎白菌肺部感染的病例分析

目的 探讨脑出血患者出现铜绿假单胞菌合并按蚊伊丽莎白菌感染的危险因素及抗菌药物治疗方案。方法 临床药师参与1例脑出血患者铜绿假单胞菌合并按蚊伊丽莎白菌肺部感染的治疗过程,通过查阅文献资料,并结合患者病情、病史、用药史及相关检查结果,分析按蚊伊丽莎白菌致病的危险因素并提出抗菌药治疗方案。结果 临床药师基于感染部位、混合细菌感染特点、抗菌药动学/药效学特性等,在抗感染方案调整为临床医师提出的用药建议后,患者全身感染得到有效控制。 结论 按蚊伊丽莎白菌为条件致病菌,毒力较低,不易感染健康人群。但当患者免疫力低时易转化为致病菌,需引起重视。     

2016-2018年清镇市第一人民医院铜绿假单胞菌的分布及耐药性分析

目的 探讨2016-2018年清镇市第一人民医院铜绿假单胞菌感染的临床分布特征及耐药性变迁,为临床合理选用抗生素治疗提供参考。方法 收集2016-2018年清镇市第一人民医院临床标本中铜绿假单胞菌的检出情况、病区分布、标本来源及药敏结果等资料,并进行统计分析。结果 共分离出铜绿假单胞菌215株,以痰液标本来源为主,占84.2%,感染病区以呼吸内科、ICU和神经外科占前3位,分别占33.9%、18.1%、17.7%。铜绿假单胞菌对复方新诺明、氨苄西林、头孢唑林、氨苄西林/舒巴坦和头孢曲松的耐药率在90.0%以上,对阿米卡星的耐药率最低。与2016年相比较,2018年铜绿假单胞菌对哌拉西林、头孢他啶、哌拉西林/他唑巴坦、头孢吡肟的耐药率呈下降趋势,对庆大霉素、妥布霉素、阿米卡星、环丙沙星、左氧氟沙星、亚胺培南的耐药率呈上升趋势。结论 2016-2018年铜绿假单胞菌对多种抗生素耐药呈上升趋势,临床应加强铜绿假单胞菌感染监控及耐药性监测,密切关注耐药性变迁,合理选用抗生素治疗。     

Early antibiotic treatment of pseudomonas aeruginosa colonisation in cystic fibrosis: a critical review of the literature

Aim To assess the evidence supporting early antibiotic treatment in asymptomatic cystic fibrosis (CF) patients colonised by Pseudomonas aeruginosa (PA).Methods We carried out a computerised (Medline, Embase) and hand search of journals for suitable publications. All English-language clinical studies regarding the efficacy of early antibiotic treatment on PA colonisation in asymptomatic patients were considered. Each eligible publications fitting these criteria were assessed for the following outcome measures: frequency of positive PA cultures; serum level of precipitating antibodies; lung function; survival; number of hospitalisations; adverse effects and resistance to antibiotics.Results Of the 11 studies eventually considered, 3 were randomised—2 versus placebo— and 8 were cohort studies—2 of which had historical controls. Overall, 309 patients (population range 7–91 patients) were recruited. There was a high variability between the individual studies for age, outcome measures, duration of follow-up (1 to 44 months) and treatment (three studies used only aerosol tobramycin, one colistin, four aerosol colistin plus oral ciprofloxacin, one used intravenous treatment and two miscellaneous therapy). An overall evaluation indicated that early antibiotic treatment can reduce the number of positive cultures and the anti-PA antibody titre. In one study, FEV1 was better in the treated group (oral ciprofloxacin and nebulised colistin) than in historical controls, while in one placebo-controlled trial, no effect on lung function was shown after 1 year of tobramycin inhalation. Collateral effects and bacterial resistance were not increased. The short follow-up did not allow definite conclusions with regard to the long-term progression of respiratory insufficiency or survival.Conclusions Evidence was found that antibiotic treatment can reduce the rate of positive cultures and of anti-PA antibody titres in asymptomatic CF patients with newly isolated PA. Different therapeutic options have not been directly compared: a multi-centre comparative study needs to be carried out.