126株铜绿假单胞菌的临床分布及耐药性分析

目的调查分析象山县中医医院铜绿假单胞菌的临床分布及药敏情况,为临床合理选用抗生素提供可靠的依据。方法采集疑似患者的标本,进行分离、培养与鉴定。采用自动微生物鉴定／药敏分析仪进行鉴定及药敏试验,对2012年7月至2013年10月分离出的126株铜绿假单胞菌（包括21株黏液型铜绿假单胞菌）的分布及耐药性进行回顾性分析。结果126株铜绿假单胞菌临床主要分布情况：痰占80．2％,尿液占11．1％,脓液占7．1％,以呼吸道感染为主。对铜绿假单胞菌保持活性较强同时耐药率〈20％的抗生素有阿米卡星、妥布霉素、亚胺培南、哌拉西林／他唑巴坦,其中碳青霉烯类耐药率升至5％,原来被认为抗铜绿假单胞菌较为有效的喹诺酮类抗生素的耐药率也有了很大提升,左氧氟沙星耐药率升至33％。黏液型铜绿假单胞菌的体外抗菌药物敏感试验耐药性较弱,且明显弱于非黏液型铜绿假单胞菌的耐药性。结论铜绿假单胞菌为医院呼吸道感染的常见致病菌,对多种抗菌药物呈不同程度耐药。加强动态监测,合理使用抗菌药物,对铜绿假单胞菌感染的预防和药物治疗具有重要指导意义。     

铜绿假单胞菌的MALDI-TOF-MS检测方法的建立

目的 建立利用基质辅助激光解吸电离飞行时间质谱仪( MALDI-TOF-MS)对铜绿假单胞菌的快速检测方法.方法 通过MALDI-TOF-MS法对铜绿假单胞菌进行检测分析,并与生化鉴定方法相比较.结果 MALDI-TOF-MS对铜绿假单胞菌的检测后得到肽指纹图片及相关质谱数据,建立MALDI-TOF-MS对铜绿假单胞菌的快速检测方法.结论 MALDI-TOF-MS方法检测铜绿假单胞菌准确快速、操作简单等特点,可发展成为食品检验铜绿假单胞菌的重要(辅助)工具.     

光声光谱技术用于绿脓杆菌的鉴定

【目的】探索一种无创、快速、可靠、经济地鉴定绿脓杆菌的新方法。【方法】利用光声光谱技术分别对大肠杆菌、绿脓杆菌在35°C下培养24 h后产生的挥发性代谢产物(Bacterial volatile compounds,BVCs)进行连续检测,获得各细菌挥发性气体的光声光谱图谱,并用“逆向思维”的方法对其检测结果进行分析。【结果】利用光声光谱技术对绿脓杆菌、大肠杆菌的挥发性代谢产物进行检测分析,发现绿脓杆菌产生了较高浓度的氰化氢(HCN),而大肠杆菌并未检测出HCN,据此可以对绿脓杆菌进行初步鉴定。【结论】光声光谱技术为绿脓杆菌的鉴定提供了一种简单、快速、经济的方法。为加速其在临床中的应用,提出了“三步走”方案:建立“大数据”、完善“比对算法”、创建“自动检测-比对-校对-鉴定”系统。     

Modified Pseudomonas agar: new differential medium for the detection/enumeration of Pseudomonas aeruginosa in mineral water.

Pseudomonas aeruginosa has been implicated as a foodborne and waterborne pathogen and is now considered a primary infectious agent. In the present study, the survival of P. aeruginosa inoculated in mineral water was evaluated by drop counts on Pseudomonas Agar Base (PAB), PAB with CN supplement X107, PAB with cetrimide, PAB with nalidixic acid, and these media with added FeSO(4). Initial counts, before starvation, were the same in all media tested. Following this period, P. aeruginosa became sensitive to PAB with added cetrimide. The addition of FeSO(4) did not improve the recovery of stressed P. aeruginosa but gave colonies a typical dark brown colour being easily differentiated from other species that can grow at 42 degrees C. The modified Pseudomonas agar medium was also tested with several P. aeruginosa strains, other species of Pseudomonas, and other genera. Only P. aeruginosa strains (pyocyanin positive) produced the typical colonies. Our results demonstrate that Pseudomonas agar with ferrous sulphate, used for the differentiation of P. aeruginosa colonies, and nalidixic acid, used as an inhibitor of Gram-positive bacteria, might be a useful medium for the detection of injured P. aeruginosa in mineral water.     

铜绿假单胞菌抗菌物质对耐甲氧西林金黄色葡萄球菌的抑菌活性研究

目的观察铜绿假单胞菌抗菌物质对耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcusaureus,MRSA)的体外抑菌活性。方法用交叉划线接种方法进行铜绿假单胞菌对32株耐甲氧西林金葡菌的体外抗菌活性的测定。结果铜绿假单胞菌抗菌物质对MRSA的体外抑菌活性良好,产生色素的菌株的抗菌活性最好,15株铜绿假单胞菌中,7株产蓝绿色色素的铜绿假单胞菌,对MRSA的抑制率均达到了100%,平均抑菌带的宽度为37.7 mm。结论铜绿假单胞菌抗菌物质对32株MRSA具有较强的抗菌活性,无疑对MRSA感染的抗菌药物研制方面开辟了一条新的途径。这是国内的首次研究报道。     

Inhibition of adhesion to buccal epithelial cells of Pseudomonas aeruginosa by dextran

Adhesion of Pseudomonas aeruginosa strains to buccal epithelial cells appears to be a necessary precondition for colonization and infection of respiratory tract. There are many strategies to prevent host organisms for Pseudomonas aeruginosa. The purpose of these studies was to evaluate the potential for preventing adhesion of Pseudomonas aeruginosa to epithelial cells with dextran. Dextran (5,000 MW) inhibited adhesion of Pseudomonas aeruginosa to buccal cells, at 20 mM was most inhibitory. The inhibitory effect appeared to be nonspecific because other neutral polysaccharides (glycogen and mannan) were also inhibitory. Dextran is an inexpansive and nontoxic agent and may be useful to prevent colonization and infection of respiratory tract with Pseudomonas aeruginosa.     

儿童ICU与普通病房铜绿假单胞菌分布及耐药性分析

目的了解铜绿假单胞菌在儿童重症监护病房（ICU）与普通病房的分布及耐药性差异,为临床合理、有效的治疗提供依据。方法分析重庆医科大学附属儿童医院2013年1月至2013年12月ICU及普通病房铜绿假单胞菌培养阳性患儿的年龄分布、菌株来源、抗菌药物的耐药性。结果 ICU和普通病房分别分离出82株和429株铜绿假单胞菌,年龄均小于2岁（70.73%和69.23%）,以痰标本来源（80.49%和64.80%）为主。ICU分离的铜绿假单胞菌对氟喹诺酮类药物的耐药率低于普通病房,对其余测试药物的耐药率均高于普通病房。ICU分离的铜绿假单胞菌中多重耐药菌55株（67.07%）,泛耐药菌44株（53.66%）,均明显高于普通病房（26.81%,3.26%）（P〈0.05）。结论铜绿假单胞菌是ICU常见的病原菌,具有多重耐药特性,加强耐药性监测,控制医院内感染,对临床医生选用有效的抗生素具有十分重要的意义。     

Lytic manifestations in the bacterial strains of Pseudomonas aeruginosa (bacteriophages bacteriocines, autoplaques).

Ps. aeruginosa strains--a frequement resuet of an irresponsible antibiotic therapy--represent a common agent of nosocomial infektions. At the same time, gravity of Pseudomonas diseases is also increasing. Lysogeny, bacteriocinogeny and frequent occurrence of autoplaques are the lytic manifestations of Pseudomonas aeruginosa strains which play a great role in the complexity of solving diagnostic, epidemiological end therapeutical problems connected with infections induced by these microbes. A survey is presented of the importance and utilization of the lytic properties of bacterial strains of Pseudomonas aeruginosa during the differentiation, epidemiological typing and further expansion of therapeutical possibilities in infections caused by Pseudomonas aeruginosa bacteria.     

Characterization by pyocine typing and serotyping of oral and sputum strains of Pseudomonas aeruginosa isolated from cystic fibrosis patients

Oral and sputum isolates of Pseudomonas aeruginosa in patients with cystic fibrosis were investigated. Of the 17 patients studied, 12 patients (71%) yielded both mucoid and nonmucoid variants of Pseudomonas aeruginosa from sputum and (or) various oral ecological sites, such as buccal mucosa, tongue dorsum, dental plaques, and saliva. A total of 51 strains of mucoid and nonmucoid Pseudomonas aeruginosa were isolated from these patients and were phenotypically characterized by both pyocine typing and serotyping. Five patients (42%) were colonized or infected by a single strain of Pseudomonas aeruginosa, whereas 7 patients (58%) were cocolonized or coinfected by two or more phenotypically different strains of Pseudomonas aeruginosa. To understand the mechanisms involved in Pseudomonas aeruginosa colonization, it may be necessary to identify multiple isolates of Pseudomonas aeruginosa not only from the sputum but also from the various oral ecological sites and to further explore the role of the oral cavity in this colonization.     

Results of using polyvalent Pseudomonas aeruginosa vaccine in children with burns by various medical centers

Polyvalent Pseudomonas aeruginosa vaccine, prepared at the Institute of Hematology from 10 hospital strains isolated from burn wounds, was administered to 32 children with extensive and deep burns. The vaccine was well tolerated. The vaccine produced a high degree of the immunity against Pseudomonas aeruginosa infection. Agglutinin serum titre increased significantly. Vaccination either prevented or inhibited the infection of burn wounds with Pseudomonas aeruginosa in all immunized children. The symptoms of Pseudomonas aeruginosa infection usually disappeared following one or two vaccinations. Bacteriemia caused by P. aeruginosa was not observed in 31 out of 32 children. In the remaining child transient bacteriemia was noted. No septicemia caused by P. aeruginosa was seen. Due to the high efficiency of the polyvalent P. aeruginosa vaccine all burned children with burns exceeding 10% of the total body surface should by vaccinated to prevent the life-threatening infections with Pseudomonas aeruginosa.     

铜绿假单胞菌生物被膜对巨噬细胞的免疫逃逸作用及机制

目的 探讨铜绿假单胞菌生物被膜对巨噬细胞的免疫逃逸作用以及相关机制。方法 用PMA刺激THP-1细胞获得巨噬细胞模型。用6孔板建膜法获得铜绿假单胞菌生物被膜菌。分别用铜绿假单胞菌的生物被膜菌和浮游菌感染巨噬细胞,观察巨噬细胞形态的变化,并检测巨噬细胞的细胞毒性和吞噬功能的变化。进一步用ELISA试剂盒检测感染的巨噬细胞培养上清中炎症细胞因子IL-1β的变化。结果 成功构建巨噬细胞模型和铜绿假单胞菌生物被膜菌模型。与感染了浮游菌的细胞相比,感染了生物被膜菌的巨噬细胞形态变化小,释放的LDH降低,吞噬功能减弱,IL-1β的表达量减少。结论 铜绿假单胞菌生物被膜菌可以逃逸巨噬细胞的免疫防御作用,其机制可能与降低巨噬细胞的炎症反应有关。     

Draft genome sequence of Pseudomonas aeruginosa strain ATCC 27853

Pseudomonas aeruginosa is a common bacterium that can cause disease. The versatility of Pseudomonas aeruginosa enables the organism to infect damaged tissues or those with reduced immunity which cause inflammation and sepsis. Here we report the genome sequence of the strain ATCC 27853.     

Isolation and Characterization of the algD gene of Pseudomonas syringae pv. phaseolicola and its distribution among other Pseudomonads and related Organisms

The gene coding for GDP-mannose dehydrogenase ( algD ) was isolated from a Pseudomonas syringae pv. phaseolicola genomic library using a polymerase chain reaction-generated heterologous DNA-probe from Pseudomonas aeruginosa . A total of 2123 base pairs were sequenced (accession number AF001555) and analysed for homologies to the alginate gene cluster of P. aeruginosa . Downstream from algD an alg8 homologue was found suggesting a similar arrangement of the alginate gene cluster in P. syringae pv. phaseolicola to that in P. aeruginosa . Also, the deduced amino acid sequence of algD shows high similarity to that of P. aeruginosa (0.9) and Azotobacter vinelandii (0.88). Southern hybridization experiments revealed that algD is widely distributed among members of the Pseudomonas rRNA homology group I. Among others, sequences homologous to algD were detected in the P. syringae pathovars lachrymans , mori , morsprunorum, pisi , savastanoi, tabaci and tomato as well as in Pseudomonas amygdali . For most of the algD positive organisms synthesis of alginate has been reported by other studies. However, algD homologues were also detected for the species Pseudomonas corrugata , Pseudomonas marginalis and Pseudomonas avenae ( Acidovorax avenae ), for which alginate biosynthesis has not yet been reported.     

Expression of the phospholipase C gene of Pseudomonas aeruginosa in Escherichia coli and Pseudomonas

The plc gene for phospholipase of Pseudomonas aeruginosa, able to be transcribed only from its own promoter, has been introduced into Escherichia coli, Pseudomonas aeruginosa and Pseudomonas putida cells in the recombinant plasmid pPMS21 of a wide host range. The expression of plc gene in all recipient cells has been shown to be phosphate regulated. The fact emphasizes the identity of pho-regulation systems in Escherichia coli and Pseudomonas cells. The level of phospholipase activity is similar in Pseudomonas putida and Pseudomonas aeruginosa under the conditions of the gene derepression, while in Escherichia coli cells the level does not exceed 10% of activity registered in Pseudomonas cells.     

不同标本来源铜绿假单胞菌体外耐药性对比研究

目的比较不同部位培养出的铜绿假单胞菌耐药性,为临床控制和治疗铜绿假单胞菌感染提供依据。方法收集住院患者不同标本中分离的铜绿假单胞菌进行鉴定和药敏试验。结果分离率以痰标本中最高,占57.67%;伤口分泌物次之占30.69%。科室分布以ICU（38.1%）、神经外科（15.34%）和烧伤科（14.81%）最多见。同一种抗菌药物对不同标本分离的铜绿假单胞菌显示出不同敏感性,痰标本中铜绿假单胞菌对常用抗菌药物耐药率较伤口分泌物标本中高,除头孢哌酮/舒巴坦、氨苄西林/舒巴坦和复方新诺明外,对其他常用的12种抗菌药物耐药率比较差异有统计学意义（P〈0.05）。结论不同标本分离的铜绿假单胞菌对抗菌药物耐药性存在差异,临床经验用药时应区别对待不同部位铜绿假单胞菌的感染。     

Synthetic peptide vaccine and antibody therapeutic development: prevention and treatment of Pseudomonas aeruginosa

Pseudomonas aeruginosa and Pseudomonas maltophilia account for 80% of opportunistic infections by pseudomonads. Pseudomonas aeruginosa is an opportunistic pathogen that causes urinary tract infections, respiratory system infections, dermatitis, soft tissue infections, bacteremia, and a variety of systemic infections, particularly in patients with severe burns, and in cancer and AIDS patients who are immunosuppressed. Pseudomonas aeruginosa is notable for its resistance to antibiotics, and is therefore a particularly dangerous pathogen. Only a few antibiotics are effective against Pseudomonas, including fluoroquinolones, gentamicin, and imipenem, and even these antibiotics are not effective against all strains. The difficulty treating Pseudomonas infections with antibiotics is most dramatically illustrated in cystic fibrosis patients, virtually all of whom eventually become infected with a strain that is so resistant that it cannot be treated. Since antibiotic therapy has proved so ineffective as a treatment, we embarked on a research program to investigate the development of a synthetic peptide consensus sequence vaccine for this pathogen. In this review article we will describe our work over the last 15 years to develop a synthetic peptide consensus sequence anti-adhesin vaccine and a related therapeutic monoclonal antibody (cross-reactive to multiple strains) to be used in the prevention and treatment of P. aeruginosa infections. Further, we describe the identification and isolation of a small peptide structural element found in P. aeruginosa strain K (PAK) bacterial pili, which has been proven to function as a host epithelial cell-surface receptor binding domain. Heterologous peptides are found in the pili of all strains of P. aeruginosa that have been sequenced to date. Several of these peptide sequences have been used in the development of an consensus sequence anti-adhesin vaccine targeted at the prevention of host cell attachment and further for the generation of a monoclonal antibody capable of prevention and treatment of existing infections.     

Transfer and integration of chromosomal genes from Pseudomonas glycinea into Pseudomonas aeruginosa.

The plasmids FP2 and R68.45 were shown to function as chromosome-mobilizing plasmids in a series of interspecific crosses between the phytopathogen Pseudomonas glycinea and the human pathogen Pseudomonas aeruginosa. At least four of seven loci tested were transferred from P. glycinea donors to P. aeruginosa auxotrophic recipients. Transductional analysis indicates that a leu+ locus of the P. glycinea chromosome transferred is stably integrated into the P. aeruginosa chromosome.     

群体感应对铜绿假单胞菌生物被膜形成的调控

生物被膜是一种与浮游细胞相对应的生长方式,由细菌和自身分泌的包外基质组成。铜绿假单胞菌是研究这一生长方式的模式生物。在过去十年,对铜绿假单胞菌生物被膜的研究已取得显著进展。群体感应(QS)的细胞沟通机制在铜绿假单胞菌生物被膜形成中发挥着重要作用。介绍生物被膜的特点,并重点讨论了QS和生物被膜之间的关系。     

健康人和感染患者抗绿脓杆菌内毒素蛋白(EP)抗体水平的测定分析

用微量间接血凝法检测了健康人和绿脓杆菌感染患者血清抗EP抗体.健康人共检测154人.基础抗体水平绝大部分在1:8以下(占85.05%)EP—HA GMT为1:7.3.无性别差异(x~2=3.04、P>0.05).绿脓杆菌感染患者检测38例,免疫前GMT高于健康人1.43倍,免疫后较免疫前GMT提高4倍.绿脓杆菌EP是绿脓杆菌共同抗原,检查人抗EP抗体水平对诊断、防治绿脓杆菌感染具有重要临床意义.     

Effects of Ambroxol on Alginate of Mature <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> Biofilms

Biofilm-forming bacteria Pseudomonas aeruginosa is a common pathogen in mechanically ventilated newborns, which can cause life-threatening infections. Alginate of mucoid Pseudomonas aeruginosa biofilms is considered an important virulence factor which contributes to the resistance to antibiotics. Traditionally, ambroxol is widely used in newborns with lung problems as a mucolytic agent and antioxidant agent as well. And there are few studies that demonstrated the anti-biofilm activity of ambroxol. In this study, we found that ambroxol can affect the structure of mucoid Pseudomonas aeruginosa biofilms. Further, we found that ambroxol reduces the production of alginate, the expression of the important genes and the activity of key enzyme guanosine diphospho-D-mannose dehydrogenase (GDP-mannose dehydrogenase; GMD) which were involved in alginate biosynthesis.