4种方法对1起铜绿假单胞菌引起感染的分型比较

目的调查外科某病区６名患者的血、大静脉插管、腹腔引流液中分离出的１４株铜绿假单胞菌医院感染。方法用生化反应分型、抗生素分型、血清型分型与ＰＣＲ分型方法对这起铜绿假单胞菌引起的医院感染进行了比较。结果生化反应分型与抗生素分型间的相关性较差，而血清型分型与ＰＣＲ分型一致。血清分型１４株菌均为ＰＡ１型；基因分型我们采用单引物ＰＣＲ指纹图分型方法，此法操作简便、快速。结论１４株铜绿假单胞菌感染是由同一血清型、同一基因型菌株所致。     

新生儿病房铜绿假单胞菌的血清学分型,质粒分析研究

新生儿病房１９８９年１０月～１９９０年１２月、１９９６年３～６月两个时间段分离４０株铜绿假单胞菌，血清学分型率９７．５％，第一时间段以Ｏ１１型最多，占３０．４％，集中在１９９０年６～１０月，第二时间段Ｏ３型占９３．８％，集中在１９９６年３～６月，表明该病房发生两次铜绿假单胞菌医院感染流行。质粒检出率４２．９％，Ｏ１１型菌株质粒谱为８．１、４６．５、１７９．１ｋｂ，并经ＨｉｎｄⅢ酶切进一步证实为同一克隆。对氟哌酸、阿米卡星及头孢他啶的敏感率分别为１００％、９７．３％和９５％。对庆大霉素耐药率明显上升，提示控制铜绿假单胞菌医院感染的重要性。     

金黄色葡萄球菌随机扩增DNA多态性分型研究

目的建立金黄色葡萄球菌随机扩增ＤＮＡ多态性（ＲＡＰＤ）分型技术，以应用于临床菌种鉴定及流行病学调查。方法对临床分离的１５株金黄色葡萄球菌，酚氯仿法提取其ＤＮＡ后行ＲＡＰＤ分型，同时进行噬菌体分型。结果１５株临床分离株大多数产生独特而稳定的ＲＡＰＤ带型，可分成５个噬菌体型和１４个ＲＡＰＤ谱型。采用同一反应体系，产肠毒素金黄色葡萄球未出现ＲＡＰＤ谱型。结论ＲＡＰＤ基因分型技术不需已知核酸序列，分型率高，分辨力强，简便快捷，可为金黄色葡萄球菌临床分离株提供分型标志，是分子流行病学研究的有效方法     

金黄色葡萄球菌随机扩增DNA多态性分型研究

目的建立金黄色葡萄球菌随机扩增ＤＮＡ多态性（ＲＡＰＤ）分型技术，以应用于临床菌种鉴定及流行病学调查。方法对临床分离的１５株金黄色葡萄球菌，酚氯仿法提取其ＤＮＡ后行ＲＡＰＤ分型，同时进行噬菌体分型。结果１５株临床分离株大多数产生独特而稳定的ＲＡＰＤ带型，可分成５个噬菌体型和１４个ＲＡＰＤ谱型。采用同一反应体系，产肠毒素金黄色葡萄球菌未出现ＲＡＰＤ谱型。结论ＲＡＰＤ基因分型技术不需已知核酸序列，分型率高，分辨力强，简便快捷，可为金黄色葡萄球菌临床分离株提供分型标志，是分子流行病学研究的有效方法。     

医院感染铜绿假单胞菌耐药谱及质粒图谱分析

目的了解医院感染铜绿假单胞菌不同血清型的耐药谱及分子流行病学特性。方法耐药谱选用Ｋ—Ｂ法和ＭＩＣ、Ａｌｋａｌｉｎｅｌｙｓｉｓ小量快速提取质粒，０．７％琼脂糖凝胶电泳获得质粒ＤＮＡ图谱。结果呼吸病区患者和氧气湿化瓶分离的ＰＡ血清４型均耐ＡＰ、ＣＺ、Ｔ和ＮＯＲ，烧伤科患者ＰＡ血清６型分别耐ＡＰ、ＣＺ、Ｔ和ＡＰ、ＣＢ、ＣＺ、ＣＸＴ、Ｔ，外科患者血清１１型均耐ＡＰ、ＣＢ、Ｐｉｐ、ＣＺ、ＣＦＰ、ＣＸＴ、ＣＡＺ、Ｔ、ＡＮ、ＧＭ和ＴＭ１１种抗生素，４株对照菌不同。质粒图谱显示：呼吸病区患者和氧气湿化瓶分离菌均有约４．０、１．８、１．７、Ｍｄａｌ３条质粒带；烧伤患者含有４．０、２．０、１．８Ｍｄａｌ３条和４．１Ｍｄａｌ１条质粒带；外科患者分离菌均有１条约３．９Ｍｄａｌ的质粒带；对照菌不同。结论采用联合分型技术对医院不同病区患者分离的不同血清型ＰＡ其同源性不同。治疗感染选用抗生素种类也不同，为临床医生在经验用药的基础上选用敏感有效抗生素起指导作用，并为医疗器械消毒效果观察提供依据。     

随机扩增多态性DNA技术用于鼠疫耶尔森氏菌基因分型的研究

目的 对来自全国不同疫源地、不同生态型的１０３株鼠疫耶尔森氏菌进行基因分型研究。方法 用随机扩增多态性ＤＮＡ技术（ＲＡＰＤ）。结果 将鼠疫耶尔森氏菌分成两种基因型别：全国大部分菌株为ＲＡＰＤ－１型,青海省境内的大部分菌株为ＲＡＰＤ－２型。结论 不同生态型的鼠疫耶尔森氏菌基因结构存在差异,为鼠疫耶尔森氏菌的基础研究和防治提供依据。     

慢性阻塞性肺病铜绿假单胞菌医院感染的菌型与防治

对７２例慢性阻塞性肺病（ＣＯＰＤ）铜绿假单胞菌感染（ＰＡ）、医院支气管肺感染（ＮＢＰＩ）的患者行痰ＰＡ血清学、噬菌体及绿脓菌素分型并做药敏试验。结果示本组ＰＡ以血清型４型为主，其绿脓菌素型为４２５３３４，依药敏选抗生素，均获良好症状控制，但仅２５％痰ＰＡ持续阴性，余反复阳性。提示ＰＡ所致ＣＯＰＤ患者ＮＢＰＩ治疗目的为控制急性症，常不能根除ＰＡ，但可有效防止新的医院感染发生。     

随机扩增多态DNA技术用于大肠杆菌基因分型的研究

采用随机扩增多态ＤＮＡ（Ｒａｎｄｏｍ ａｍｐｌｉｆｉｅｄ ｐｏｌｙｍｏｐｈｉｃ ＤＮＡ，简称ＲＡＰＤ）技术，用二组引物对８株大肠杆菌进行了基因分型，发现二组引物均能将８株菌分成４个基因型，其中血清型相同，但来自不同国家和地区的３株被分成二个基因型，表明这三株菌基因组ＤＮＡ间存在差异。４株从同一病人体内不同次分离到的血清型相同的菌株呈同一指纹图谱。说明该４株菌渊源于同一菌株，结果显示该方法的可分型性     

老年肺部感染患者铜绿假单胞菌血清学分型及对抗生素体外抗菌活性研究

对老年肺部感染患者呼吸道分泌物分离的６８株铜绿假单胞菌进行了分型鉴定。６８株铜绿假单菌以６、２、１１、１９和１型为主，占分型菌株的７３．５％，同时我们还进行了铜绿假胞单菌１５种抗生素的敏感性测定。老年肺部感染患者铜绿假单胞菌对悉复欢的敏感率为８６．７６％、对多粘菌素Ｂ的敏感率为８３．８％、对氨苄青霉素的耐药率最高９７．１％、对其它１１种抗生素有不同的敏感性。本结果对老年肺部感染患者铜绿假单胞菌的流行病学调查及铜绿假单胞菌感染临床治疗的药物选择具有重要意义。     

老年肺部感染患者铜绿假单胞菌血清学分型及对抗生素体…

对老年肺部感染患者呼吸道分泌物分离的６８株铜绿假单胞菌进行了分型鉴定。６８株铜绿假单菌以６、２、１１、１９和１型为主，占分型力株的７３．５％同时我们还进行了铜绿假胞单菌１５种抗生素的敏感性测定。老年肺部感染患者铜绿假单胞菌和欢的敏感率为８５６．７６％、对多粘菌素Ｂ的敏感８３．８％，对氨苄青霉寒的药率最高９７．１％，对其它１１种抗生素有不同的敏感性，本结果对老年肺部感染患者铜绿单细胞胞菌的流行病学调     

Molecular characterization of Vibrio cholerae O1 and non-O1 from human and environmental sources in Malaysia

A total of 31 strains of Vibrio cholerae O1 (10 from outbreak cases and 7 from surface water) and non-O1 (4 from clinical and 10 from surface water sources) isolated between 1993 and 1997 were examined with respect to presence of cholera enterotoxin (CT) gene by PCR-based assays, resistance to antibiotics, plasmid profiles and random amplified polymorphic DNA (RAPD) analysis. All were resistant to 9 or more of the 17 antibiotics tested. Identical antibiotic resistance patterns of the isolates may indicate that they share a common mode of developing antibiotic resistance. Furthermore, the multiple antibiotic resistance indexing showed that all strains tested originated from high risk contamination. Plasmid profile analysis by agarose gel electrophoresis showed the presence of small plasmids in 12 (7 non-O1 and 5 O1 serotypes) with sizes ranging 1.3-4.6 MDa. The CT gene was detected in all clinical isolates but was present in only 14 (6 O1 serotype and 8 non-O1 serotype) isolates from environmental waters. The genetic relatedness of the clinical and environmental Vibrio cholerae O1 and non-O1 strains was investigated by RAPD fingerprinting with four primers. The four primers generated polymorphisms in all 31 strains of Vibrio cholerae tested, producing bands ranging from < 250 to 4500 bp. The RAPD profiles revealed a wide variability and no correlation with the source of isolation. This study provides evidence that Vibrio cholerae O1 and non-O1 have significant public health implications.     

ICU铜绿假单胞菌感染暴发的随机扩增多态性DNA分子分型 FREE

目的对铜绿假单胞菌（PA）进行随机扩增多态性ＤＮＡ(RAPD)分子分型,探讨重症监护室(ICU)中PＡ医院感染的流行规律。方法采用RAPD技术对13株分离自某院ICU临床诊断为医院感染肺炎患者下呼吸道标本的PＡ进行分型，并与抗菌药物耐药谱分型比较。结果13株PＡ产生A型（12株，92.31%）和B型（1株，7.69%）2种耐药表型，优势耐药表型为A型；RAPD分型可分2型，分别为Ⅰ型(6株，46.15%)和Ⅱ型(7株，53.85%)。7株分子Ⅱ型和5株分子Ⅰ型的耐药A型PA是优势菌株，可判断为引起ICU中PA医院感染暴发的病原菌。结论ICU存在PA暴发流行，流行株为耐药A型/分子Ⅱ型、耐药A型/分子Ⅰ型。PA产生多重耐药性，RAPD分型是目前比较理想的分子流行病学溯源手段。     

Vegetables as a source of infection with Pseudomonas aeruginosa in a University and Oncology Hospital of Rio de Janeiro.

Samples of fresh vegetables fed to patients in an Oncology and a University Hospital were examined for frequency of recovery and counts of Pseudomonas aeruginosa. Thirty-eight isolates from vegetables as well as 98 clinical isolates recovered during the same period of vegetable collection were serotyped and assayed for pyocin production in order to evaluate the role of vegetables as a source of microorganisms. Pseudomonas aeruginosa was recovered from 19.0% of the vegetable samples. Although 1% hypochlorite solution was used as a sanitizer, 50% of the positive samples were found to harbour more than 100 colony-forming units (cfu) g-1. Lettuce, chicory and watercress yielded the highest frequencies of isolation (P less than 0.05). The pyocin typing and serotyping of clinical strains revealed some types identical to those recovered from vegetables. Among those found in the University Hospital, serotype O4 and pyocin type PT10/b were detected in vegetables and in clinical specimens whereas types O1-PT22/e, O2a-PT10/a, O2a-PT10/b, O4-PT10/a, O11-PT10/a and O11-PT10/b were common in both groups of strains isolated in the Oncology Hospital. Our results strongly suggest that vegetables represent a source of endemic infection with P. aeruginosa for hospitalized patients.     

Multiresistant serotype O 12 Pseudomonas aeruginosa: evidence for a common strain in Europe

A survey was made of serotype association and multiple antibiotic resistance in strains of Pseudomonas aeruginosa in Europe. Of 208 epidemiologically distinct strains from 16 laboratories in 10 countries, 48 were resistant to carbenicillin (MIC greater than 128 micrograms/ml) and gentamicin (MIC greater than 4 micrograms/ml), and 12 of these strains were of serotype O 12. Representative O 12 strains from different countries were compared with two multiresistant O 12 strains isolated 5 years apart, from a British burns unit and the antibiotic sensitive serotype reference strain. All O 12 strains were similar by phage and pyocin typing but lysogenic phage profiles indicated that two strains (the later burns isolate and the serotype strain) were distinct from the others. Electrophoretic characterization of outer membrane proteins, lipopolysaccharides and esterase enzymes corroborated the relationship of the strains and restriction fragment length polymorphism of DNA fragments hybridized with a cDNA probe copy of rRNA from P. aeruginosa provided further proof of their relatedness. We propose that the uniformity of characters of multiresistant O 12 P. aeruginosa in Europe is suggestive of a common origin for the strains.     

重症监护室内鲍曼不动杆菌随机扩增DNA多态性分型研究

目的：建立鲍曼不动杆菌随机扩增DNA多态性（RAPD）分型技术,以用于临床菌种鉴定及流行病学调查。方法：收集6个月内从ICU内分离出的14株鲍曼不动杆菌,提取DNA后进行RAPD分型;同时进行生物学分型和抗生素敏感性实验,结果：14株菌株被分为5种RAPD型,其中有5株属于同一种类型（A型）,有6株属于另外同一种类型（B型）,其他3株分别属于另3种不同类型（C,D,E型）,而生物学分型只有两种类型（生物型1,9）,抗生素敏感性实验表明14株菌株均为仅对亚胺培南／西司他丁（泰能）,头孢哌酮/舒巴坦（舒普深）等少数几种抗生素敏感的多重耐药菌株。结论：ICU内存在鲍曼不动杆菌暴发流行,RAPD分型技术分型率高,分辨力强,简便快速,具有分子流行病学研究的应用价值。     

Phenotypic stress response of Pseudomonas aeruginosa following culture in water microcosms

The purpose of the present study was to explore the potential behavioural changes of Pseudomonas aeruginosa following growth in different aquatic environmental conditions. To achieve this, P. aeruginosa was cultured in various water microcosms for 12 months under fixed (pH, nutrients and temperature) factors. P. aeruginosa responses to these conditions were investigated using colony morphotype, biochemical and enzymatic characterisation, pyocin typing, serotyping, sensitivity to different classes of antibiotics and molecular identification. Results show that starvation in water microcosms lead to unusual phenotypes. Of interest is that the pyocin changed from 24/n in the wild type to 83/a following culture in the water microcosms, and the serotype changed from O6 in the wild type to O1 in microcosm-cultured P. aeruginosa. Furthermore, the starvation period in various aquatic microcosms enhanced the resistance of P. aeruginosa against beta-lactam antibiotics. Compared to the other aquatic environments, the seawater microcosm produced the greatest amount of variations in P. aeruginosa. Overall, data demonstrated a high adaptability of P. aeruginosa to environmental changes. This may explain the unusual antibiotic-resistant phenotypes belonging to P. aeruginosa species, and their capacity for spreading that leads to human infections.     

Bacterial contamination of dental chair units in a modern dental hospital caused by leakage from suction system hoses containing extensive biofilm

Within six months of opening of the new Dublin Dental Hospital in September 1998, areas of corrosion were observed on many of the baseplates of the hospital's 103 dental chair units (DCUs) at the site of attachment of the suction hoses. The corroded areas were heavily contaminated with Pseudomonas spp. and related genera posing a risk of cross-infection, particularly for immunocompromised patients. These species were used as marker organisms to investigate the source of the contamination. P. aeruginosa was the predominant species recovered from 41 selected DCU baseplates (61% prevalence), whereas P. putida (46% prevalence) and P. aeruginosa (43% prevalence) were predominant at the attachment ends of 37 selected high-volume suction hoses. Forty-one selected isolates of P. aeruginosa from 13 DCU baseplates, 16 high-volume suction hoses and 12 coarse filter housings (another suction system site) from 19 separate DCUs were serotyped to determine the similarity of isolates at each site. The majority of isolates (68.3%) belonged to serotype O:10, while the remainder belonged to serotypes O:6 (7.3%), O:11 (7.3%), O:14 (9.8%) and O:5/O:16 (7.3%). Of the isolates from DCU baseplates, additional isolates with the same serotype were recovered from other suction system sites in 10/13 (77%) cases. Isolates of only one serotype were recovered from each of the 19 DCUs investigated. Forty-one serotyped isolates were also subject to computer-assisted analysis of SpeI-generated DNA fingerprint profiles, and similarity coefficient (S(AB)s) values were calculated for each pairwise combination of isolate profiles. The data obtained showed that the isolates consisted of two distinct main populations, each containing separate clades corresponding to specific serotypes. Serotype O:6 (three isolates), O:11 (three isolates) and O:5/O:16 (three isolates) belonged to a single strain in each case. Serotypes O:14 (four isolates) and O:10 (28 isolates) belonged to two strains in each case. The two serotype O:10 strains, termed fingerprint groups I (four isolates from three DCUs) and II (24 isolates from 10 DCUs), were the most distantly related of all the strains identified. These findings demonstrated that the hospital DCUs had become colonized with a small number of P. aeruginosa strains, one of which (serotype O:10, fingerprint group II) predominated. These results also confirmed that DCU baseplate contamination was most likely to be due to leakage from suction system hoses at the baseplate attachment sites, probably due to loosening during use. Replacement hose connectors that firmly retained the suction hoses in the attachment sites so that they could not be loosened by movement of the suction hoses solved this problem, and eliminated further contamination of the DCU baseplates.     

Pseudomonas aeruginosa isolated in immunocompromised patients: antimicrobial resistance, serotyping, and molecular typing

OBJECTIVE: Our study dealt with antibiotic resistance and serotypes of Pseudomonas aeruginosa strains isolated from immunocompromised patients in the National Bone Marrow Transplant Center of Tunis as well as molecular typing of ceftazidime resistant strains (CAZ-R). DESIGN: We studied a total of 87 non-replicate P. aeruginosa isolates from 36 patients (84 strains) or the hospital environment (3 strains). RESULTS: Rates of antimicrobial resistance were 36% for ceftazidime, 16% for imipenem, 38% for amikacin, and 57% for ciprofloxacin. The 31 CAZ-R strains were associated with O:11 serotype in 84% of the cases. Genetically characterization of CAZ-R strains by Pulsed Field Gel Electrophoresis (PFGE) after digestion of genomic DNA with SpeI revealed 2 genotypic groups. The first was composed of strains isolated from one outpatient between November 1998 and April 1999. Resistance phenotypes of these strains varied after use of antimicrobial drugs. The second was predominant (18/31 CAZ-R strains) in both hematology and graft units and persisted from June 1998 to June 2000 among 5/8 patients. These strains had O:11 serotype in 78% of the cases. The strains of this group were not isolated on patient admission and were isolated from 2 washbasins in the graft unit in May 1999. CONCLUSION: These results suggest the spread of multidrug-resistant O:11 P. aeruginosa clone from a tap water among hospitalized patients in our center, emphasizing the need of standard control of washbasins to eradicate this reservoir.     

Identification of infectious Pseudomonas aeruginosa strains in an occupational saturation diving environment

OBJECTIVES: Occupational saturation divers have various skin disorders, of which skin infections are the most serious and frequent. Pseudomonas aeruginosa is the microbe most often isolated from skin infections in divers. The purpose of the present work was (a) to report the occurrence of P aeruginosa in skin infections in operational saturation diving in the North Sea from 1987 to 1995; (b) to report the environmental occurrence of P aeruginosa in saturation diving systems, and finally (c) to identify possible relations between infection related to strains of P aeruginosa and environmental isolates of the microbe. RESULTS: During the period 1987-95, P aeruginosa was isolated from 257 skin infections in operational saturation divers. Most of the isolates related to infection by P aeruginosa show a unique growth inhibition pattern towards the normal skin flora, and the serotype pattern of P aeruginosa from skin infections is limited compared with similar infections in non-divers. In a mini-epidemiological study on board one diving vessel during one operational diving period, five significantly different DNA fragment profiles were found among the 12 isolates related to infection by P aeruginosa obtained from the saturation system. In two cases the infectious genotypes were detected in the fresh water for the saturation chambers weeks before the arrival of the infected diver. CONCLUSIONS: The most commonly used epidemiological marker for P aeruginosa world wide, also used in earlier studies, is serotyping, but with pulsed field gel electrophoresis (PFGE) miniepidemiology it was shown to be insufficient for epidemiological purposes in saturation environments. PFGE analyses were shown to be superior both to antibacterial factor and to serotyping in epidemiological analyses of P aeruginosa infections in saturation diving.     

霍乱疫情分离株流行菌型及耐药性分析

目的了解霍乱疫情分离株的流行菌型及其耐药性,为霍乱疫情的快速有效控制和临床治疗提供科学依据。方法对2008年6月海南省某地霍乱疫情采集的6株霍乱弧菌分离株进行血清学分型、噬菌体-生物分型、霍乱毒素基因检测、脉冲场凝胶电泳(PFGE)分子分型和药物敏感性试验。结果 6株菌均属埃尔托生物型霍乱弧菌,除1株为O1群小川型霍乱弧菌流行株(1c),其余5株均为O1群稻叶型霍乱弧菌流行株(1c);6株菌均具有霍乱毒素基因;PFGE分型为2个型,但聚类图谱分析相似性为100%;6株菌均对链霉素、复方新诺明、磺胺异噁唑、多粘菌素B耐药;对诺氟沙星、头孢噻肟、头孢噻吩等9种抗生素均敏感;结论该起霍乱疫情的流行菌型为O1群埃尔托霍乱弧菌稻叶型流行株(1c);诺氟沙星、头孢噻肟、头孢噻吩等可作为病例和带菌者治疗的指导用药,链霉素、复方新诺明、磺胺异噁唑、多粘菌素B则不宜使用。